Utilización del cultivo plurianual de pataca (Helianthus tuberosus L.) para la producción de hidratos de carbono fermentables a partir de los tallos

La pataca (Helianthus tuberosus L.) es una especie de cultivo con un alto potencial en la producción de hidratos de carbono de reserva en forma de polifructanos, especialmente inulina, que se acumulan temporalmente en los tallos en forma de polisacáridos para translocarse posteriormente a los tubérculos, donde son almacenados. Aunque tradicionalmente el producto de interés del cultivo son los tubérculos, que acumulan gran cantidad de hidratos de carbono fermentables (HCF) cuando se recogen al final del ciclo de desarrollo, en este trabajo se pretende evaluar el potencial de la pataca como productor de HCF a partir de los tallos cosechados en el momento de máximo contenido en HCF, mediante un sistema de cultivo plurianual. Se han realizado los siguientes estudios: i) Determinación del momento óptimo de cosecha en ensayos con 12 clones ii) Potencial del cultivo plurianual de la pataca en términos de producción anual de biomasa aérea y de HCF en cosechas sucesivas, iii) Ensayos de conservación de la biomasa aérea, iv) Estimación de los costes de las dos modalidades de cultivo de pataca para producción de HCF y v) Estimación de la sostenibilidad energética de la producción de bioetanol mediante la utilización de los subproductos. Para la determinación del momento óptimo de la cosecha de la biomasa aérea se ensayaron 12 clones de diferente precocidad en Madrid; 4 tempranos (Huertos de Moya, C-17, Columbia y D-19) y 8 tardíos (Boniches, China, K-8, Salmantina, Nahodka, C-13, INIA y Violeta de Rennes). El máximo contenido en HCF tuvo lugar en el estado fenológico de botón floral-flor que además coincidió con la máxima producción de biomasa aérea. De acuerdo con los resultados obtenidos, la cosecha de los clones tempranos se debería realizar en el mes de julio y en los clones tardíos en septiembre, siendo éstos últimos más productivos. La producción media más representativa entre los 12 clones, obtenida en el estado fenológico de botón floral fue de 23,40 t ms/ha (clon INIA), con un contenido medio en HCF de 30,30 % lo que supondría una producción potencial media de 7,06 t HCF/ha. La producción máxima en HCF se obtuvo en el clon Boniches con 7,61 t/ha y 22,81 t ms/ha de biomasa aérea. En el sistema de cultivo plurianual la cantidad de tallos por unidad de superficie aumenta cada año debido a la cantidad de tubérculos que van quedando en el terreno, sobre todo a partir del 3er año, lo que produce la disminución del peso unitario de los tallos, con el consiguiente riesgo de encamado. El aclareo de los tallos nacidos a principios de primavera mediante herbicidas tipo Glifosato o mediante una labor de rotocultor rebaja la densidad final de tallos y mejora los rendimientos del cultivo. En las experiencias de conservación de la biomasa aérea se obtuvo una buena conservación por un período de 6 meses de los HCF contenidos en los tallos secos empacados y almacenados bajo cubierta. Considerando que el rendimiento práctico de la fermentación alcohólica es de 0,5 l de etanol por cada kg de azúcar, la producción potencial de etanol para una cosecha de tallos de 7,06 t de HCF/ha sería de 3.530 l/ha. El bagazo producido en la extracción de los HCF de la biomasa aérea supondría 11,91 t/ha lo que utilizado para fines térmicos supone más de 3 veces la energía primaria requerida en el proceso de producción de etanol, considerando un poder calorífico inferior de 3.832,6 kcal/kg. Para una producción de HCF a partir de la biomasa aérea de 7,06 t/ha y en tubérculos al final del ciclo de 12,11 t/ha, los costes de producción estimados para cada uno de ellos fueron de 184,69 €/t para los HCF procedentes de la biomasa aérea y 311,30 €/t para los de tubérculos. Como resultado de este trabajo se puede concluir que la producción de HCF a partir de la biomasa aérea de pataca en cultivo plurianual, es viable desde un punto de vista técnico, con reducción de los costes de producción respecto al sistema tradicional de cosecha de tubérculos. Entre las ventajas técnicas de esta modalidad de cultivo, cabe destacar: la reducción de operaciones de cultivo, la facilidad y menor coste de la cosecha, y la posibilidad de conservación de los HCF en la biomasa cosechada sin mermas durante varios meses. Estas ventajas, compensan con creces el menor rendimiento por unidad de superficie que se obtiene con este sistema de cultivo frente al de cosecha de los tubérculos. Jerusalem artichoke (Helianthus tuberosus L.) (JA) is a crop with a high potential for the production of carbohydrates in the form of polyfructans, especially inulin, which are temporarily accumulated in the stems in the form of polysaccharides. Subsequently they are translocated to the tubers, where they are finally accumulated. In this work the potential of Jerusalem artichoke for fermentable carbohydrates from stems that are harvested at their peak of carbohydrates accumulation is assessed as compared to the traditional cultivation system that aims at the production of tubers harvested at the end of the growth cycle. Tubers are storage organs of polyfructans, namely fermentable carbohydrates. Studies addressed in this work were: i) Determination of the optimum period of time for stem harvesting as a function of clone precocity in a 12-clone field experiment; ii) Study of the potential of JA poly-annual crop regarding the annual yield of aerial biomass and fermentable carbohydrates (HCF) over the years; iii) Tests of storage of the aerial biomass, iv) Comparative analysis of the two JA cultivation systems for HCF production: the poly-annual system for aerial biomass harvesting versus the annual cultivation system for tubers and v) Estimation of the energy sustainability of the bioethanol production by using by-products of the production chain. In order to determine the best period of time for aerial biomass harvesting twelve JA clones of different precocity were tested in Madrid: four early clones (Huertos de Moya, C-17, Columbia and D-19) and eight late clones (Boniches, China, K-8 , Salmantina, Nahodka, C-13, INIA and Violeta de Rennes). Best time was between the phenological stages of floral buds (closed capitula) and blossom (opened capitula), period in which the peak of biomass production coincides with the peak of HCF accumulation in the stems. According to the results, the early clones should be harvested in July and the late ones in September, being the late clones more productive. The clone named INIA was the one that exhibited more steady yields in biomass over the 12 clones experimented. The average potential biomass production of this clone was 23.40 t dm/ha when harvested at the floral buds phenological stage; mean HCF content is 30.30%, representing 7.06 t HCF/ha yield. However, the highest HCF production was obtained for the clone Boniches, 7.61 t HCF/ha from a production of 22.81 t aerial biomass/ha. In the poly-annual cultivation system the number of stems per unit area increases over the years due to the increase in the number of tubers that are left under ground; this effect is particularly important after the 3rd year of the poly-annual crop and results in a decrease of the stems unit weight and a risk of lodging. Thinning of JA shoots in early spring, by means of an herbicide treatment based on glyphosate or by means of one pass with a rotary tiller, results in a decrease of the crop stem density and in higher crop yields. Tests of biomass storing showed that the method of keeping dried stems packed and stored under cover results in a good preservation of HCF for a period of six months at least. Assuming that the fermentation yield is 0.5 L ethanol per kg sugars and a HCF stem production of 7.06 t HCF/ha, the potential for bioethanol is estimated at 3530 L/ha. The use of bagasse -by-product of the process of HCF extraction from the JA stems- for thermal purposes would represent over 3 times the primary energy required for the industrial ethanol production process, assuming 11.91 t/ha bagasse and 3832.6 kcal/kg heating value. HCF production costs of 7.06 t HCF/ha yield from aerial biomass and HCF production costs of 12.11 t HCF/ha from tubers were estimated at 184.69 €/t HCF and 311.30 €/t HCF, respectively. It can be concluded that the production of HCF from JA stems, following a poly-annual cultivation system, can be feasible from a technical standpoint and lead to lower production costs as compared to the traditional annual cultivation system for the production of HCF from tubers. Among the technical advantages of the poly-annual cultivation system it is worth mentioning the reduction in crop operations, the ease and efficiency of harvesting operations and the possibility of HCF preservation without incurring in HCF losses during the storage period, which can last several months. These advantages might compensate the lower yield of HCF per unit area that is obtained in the poly-annual crop system, which aims at stems harvesting, versus the annual one, which involves tubers harvesting.

Localización óptima de una planta de bioetanol a partir de tubérculos de pataca (Helianthus tuberosus l.) en la Cuenca del Duero

El objetivo del presente trabajo es determinar la localización óptima de una planta de producción de 30.000 m3/año de bioetanol a partir de tubérculos de pataca (Helianthus tuberosus L.) cultivada en regadío, en tierras de barbecho de la Cuenca Hidrográfica del Duero (CH Duero). Inicialmente se elaboró, a partir de datos bibliográficos, un modelo de producción de pataca en base a una ecuación de regresión que relaciona datos experimentales de rendimientos de variedades tardías con variables agroclimáticas. Así se obtuvo una función de producción basada en la cantidad de agua disponible (precipitación efectiva + dosis de riego) y en la radiación global acumulada en el periodo brotación‐senescencia del cultivo. A continuación se estima la superficie potencial de cultivo de pataca en la CH Duero a partir de la superficie arable en regadío cartografiada por el Sistema de Ocupación del Suelo (SIOSE), a la cual se le aplican, en base a los requerimientos del cultivo, unas restricciones climáticas, edafológicas, topográficas y logísticas mediante el uso de Sistemas de Información Geográfica (SIG). La proporción de superficie de regadío restringida se cuantifica a escala municipal con el fin de calcular la superficie de barbecho en regadío apta para el cultivo de pataca. A partir de las bases de datos georreferenciadas de precipitación, radiación global, y la dotación de agua para el riego de cultivos no específicos establecida en el Plan Hidrológico de la Cuenca del Duero a escala comarcal, se estimó la producción potencial de tubérculos de pataca sobre la superficie de barbecho de regadío según el modelo de producción elaborado. Así, en las 53.360 ha de barbecho en regadío aptas para el cultivo de pataca se podrían producir 3,8 Mt de tubérculos al año (80 % de humedad) (761.156 t ms/año) de los que se podría obtener 304.462 m3/año de bioetanol, considerando un rendimiento en la transformación de 12,5 kg mf/l de etanol. Se estiman los costes de las labores de cultivo de pataca así como los costes de la logística de suministro a una planta de transformación considerando una distancia media de transporte de 25 km, en base a las hojas de cálculo de utilización de aperos y maquinaria agrícola oficiales del Ministerio de Agricultura, Alimentación y Medio Ambiente (MAGRAMA). Considerando el balance de costes asociados a la producción de bioetanol (costes de transformación, distribución y transporte del producto, costes estructurales de la planta, ahorro de costes por la utilización de las vinazas generadas en el proceso como fertilizante y un beneficio industrial), se ha estimado que el coste de producción de bioetanol a partir de tubérculos de pataca asciende a 61,03 c€/l. Se calculan los beneficios fiscales para el Estado por el cultivo de 5.522 ha de pataca que suministren la materia prima necesaria para una planta de bioetanol de 30.000 m3/año, en concepto de cotizaciones a la Seguridad Social de los trabajadores, impuestos sobre el valor añadido de los productos consumidos, impuesto sobre sociedades y ahorro de las prestaciones por desempleo. Se obtuvieron unos beneficios fiscales de 10,25 c€ por litro de bioetanol producido. El coste de producción de bioetanol depende del rendimiento de tubérculos por hectárea y de la distancia de transporte desde las zonas de producción de la materia prima hasta la planta. Se calculó la distancia máxima de transporte para que el precio de coste del bioetanol producido sea competitivo con el precio de mercado del bioetanol. Como resultado se determinó que el precio del bioetanol (incluido un beneficio industrial del 15%) de la planta sería igual o inferior al precio de venta en el mercado (66,35 c€/l) con una distancia máxima de transporte de 25 km y un rendimiento mínimo del cultivo de 60,1 t mf/ha. Una vez conocido el área de influencia de la planta según la distancia de transporte máxima, se determinó la localización óptima de la planta de producción de bioetanol mediante un proceso de ubicación‐asignación realizado con SIG. Para ello se analizan los puntos candidatos a la ubicación de la planta según el cumplimiento de unos requerimientos técnicos establecidos (distancia a fuentes de suministro eléctrico y de recursos hídricos, distancia a estaciones de ferrocarril, distancia a núcleos urbanos y existencia de Espacios Naturales Protegidos) que minimizan la distancia de transporte maximizando la cantidad de biomasa disponible según la producción potencial estimada anteriormente. Por último, la superficie destinada al cultivo de pataca en el área de influencia de la planta se determina en base a un patrón de distribución del cultivo alrededor de una agroindustria. Dicho patrón se ha obtenido a partir del análisis del grado de ocupación del cultivo de la remolacha en función de la distancia de transporte a la planta azucarera de Miranda de Ebro (Burgos). El patrón resultante muestra que la relación entre el grado de ocupación del suelo por el cultivo y la distancia de transporte a la planta siguen una ecuación logística. La localización óptima que se ha obtenido mediante la metodología descrita se ubica en el municipio leonés de El Burgo Ranero, donde la producción potencial de tubérculos de pataca en la superficie de barbecho situada en un radio de acción de 25 km es de 375.665 t mf/año, superando las 375.000 t mf requeridas anualmente por la planta de bioetanol. ABSTRACT Jerusalem artichoke (Helianthus tuberosus L.) is a harsh crop with a high potential for biomass production. Its main use is related to bioethanol production from the carbohydrates, inulin mainly, accumulated in its tubers at the end of the crop cycle. The aerial biomass could be used as solid biofuel to provide energy to the bioethanol production process. Therefore, Jerusalem artichoke is a promising crop as feedstock for biofuel production in order to achieve the biofuels consumption objectives established by the Government of Spain (PER 2011‐2020 and RDL 4/2013) and the European Union (Directive 2009/28/EC). This work aims at the determination of the optimal location for a 30,000 m3/year bioethanol production plant from Jerusalem artichoke tubers in the Duero river basin. With this purpose, a crop production model was developed by means of a regression equation that relates experimental yield data of late Jerusalem artichoke varieties with pedo‐climatic parameters from a bibliographic data matrix. The resulting crop production model was based on the crop water availability (including effective rainfall and irrigation water supplied) and on global radiation accumulated in the crop emergence‐senescence period. The crop potential cultivation area for Jerusalem artichoke in the Duero basin was estimated using the georeferenced irrigated arable land from the “Sistema de Ocupación del Suelo” (SIOSE) of Spain. Climatic, soil, slope and logistic restrictions were considered by means of Geographic Information Systems (GIS). The limited potential growing area was then applied to a municipality scale in order to calculate the amount of fallow land suitable for Jerusalem artichoke production. Rainfall and global radiation georeferenced layers as well as data of irrigation water supply for crop production (established within the Duero Hydrologic Plan) were use to estimate the potential production of Jerusalem artichoke tubers in the suitable fallow land according to the crop production model. As a result of this estimation, there are 53,360 ha of fallow land suitable for Jerusalem artichoke production in the Duero basin, where 3.8 M t fm/year could be produced. Considering a bioethanol processing yield of 12.5 kg mf per liter of bioethanol, the above mentioned tuber potential production could be processed in 304,462 m3/year of bioethanol. The Jerusalem crop production costs and the logistic supply costs (considering an average transport distance of 25 km) were estimated according to official agricultural machinery cost calculation sheets of the Minister of Agriculture of Spain (MAGRAMA). The bioethanol production cost from Jerusalem artichoke tubers was calculated considering bioethanol processing, transport and structural costs, industrial profits as well as plant cost savings from the use of vinasses as fertilizer. The resulting bioetanol production cost from Jerusalem artichoke tubers was 61.03 c€/l. Additionally, revenues for the state coffers regarding Social Security contributions, added value taxes of consumed raw materials, corporation tax and unemployment benefit savings due to the cultivation of 5,522 ha of Jerusalem artichoke for the 30.000 m3/year bioethanol plant supply were calculated. The calculated revenues amounted to 10.25 c€/l. Bioethanol production cost and consequently the bioethanol plant economic viability are strongly related to the crop yield as well as to road transport distance from feedstock production areas to the processing plant. The previously estimated bioethanol production cost was compared to the bioethanol market price in order to determine the maximum supply transport distance and the minimum crop yield to reach the bioethanol plant economic viability. The results showed that the proposed plant would be economically viable at a maximum transport distance of 25 km and at a crop yield not less than 60.1 t fm/ha. By means of a GIS location‐allocation analysis, the optimal bioethanol plant location was determined. Suitable candidates were detected according to several plant technical requirements (distance to power and water supply sources, distance to freight station, and distance to urban areas and to Natural Protected Areas). The optimal bioethanol plant location must minimize the supply transport distance whereas it maximizes the amount of available biomass according to the previously estimated biomass potential production. Lastly, the agricultural area around the bioethanol plant finally dedicated to Jerusalem artichoke cultivation was planned according to a crop distribution model. The crop distribution model was established from the analysis of the relation between the sugar beet (Beta vulgaris L.) cropping area and the road transport distance from the sugar processing plant of Miranda de Ebro (Burgos, North of Spain). The optimal location was situated in the municipality of ‘El Burgo Ranero’ in the province of León. The potential production of Jerusalem artichoke tubers in the fallow land within 25 km distance from the plant location was 375,665 t fm/year, which exceeds the amount of biomass yearly required by the bioethanol plant.

我国严重威胁性外来入侵植物入侵与扩散历史过程重建及其潜在分布区的预测

外来生物入侵已经在世界范围内造成严重的危害，它不仅导致生物多样性的减少和丧失而且威胁着全球的生态环境和经济发展。入侵种在入侵区域的种群扩散是入侵种带来风险的最根本问题，也是其造成危害的重要原因。研究入侵物种的扩散规律可以了解其在入侵地的入侵状况和扩散趋势，这对制定合理的控制措施具有重要意义。本研究选取14种在我国具有较强威胁性的外来入侵植物，通过对其入侵历史动态的分析，判断其在我国的入侵与扩散阶段，进而预测其在我国的潜在分布区。这14种外来植物为紫茎泽兰（Ageratina adenophora）、土荆芥 （Chenopodium ambrosioides.）、喜旱莲子草（Alternanthera philoxeroides）、反枝苋（Amaranthus retroflexus）、刺苋（Amaranthus spinosus）、皱果苋（Amaranthus viridis）、北美独行菜（Lepidium virginicum）、藿香蓟（Ageratum conyzoides）、钻形紫菀（Aster subulatus）、小蓬草（Conyza canadensis）、一年蓬（Erigeron annuus）、牛膝菊（Galinsoga paviflora）、飞机草（Eupatorium odorata）和北美商陆（Phytolacca americana）。 　　紫茎泽兰最早于20世纪40年代入侵我国，经过1940—1960年长达20年的时滞期，紫茎泽兰开始在云南及其临近的省份如四川、贵州和广西迅速扩散。其中，紫茎泽兰在南亚热带和中亚热带气候条件下的扩散速度为20公里/年，而在垂直地带性北亚热带地区的扩散速度为6.8公里/年。紫茎泽兰仍没有入侵到垂直地带性暖温带地区。尽管1990年后，紫茎泽兰在云南基本停止扩散，但其在邻近省（市）的快速传播表明紫茎泽兰在我国仍处在扩散阶段，还没有达到饱和阶段。生态位模型预测结果和紫茎泽兰在已经入侵地区扩散的地理生态式样基本相符。我国南部及中南部地区的气候条件十分适合紫茎泽兰生长。而在华中地区，由于环境条件不太适宜，其扩散速度会相对较慢。我国北部和西北部的气候条件完全不适合紫茎泽兰生存。因此，我国南部和中南部广大未入侵地区将会受到紫茎泽兰入侵和快速扩散的严重威胁，对此应该立即采取紧急措施。 　　自从紫茎泽兰于1978年首次入侵四川省以来，已经在四川、重庆和湖北大面积扩散。紫茎泽兰在此新近入侵区域主要沿河流、公路、铁路扩散。其中沿安宁河、108国道和成昆铁路向北扩散的平均速度为19公里/年，而沿金沙江－长江向东北扩散的平均速度为33公里/年。特别是2000年以后，紫茎泽兰沿金沙江－长江的扩散速度达到了88公里/年。紫茎泽兰得以如此迅速扩散是其本身生物学特性和当地地理生态特点多种因素之间一系列耦合关系共同作用的结果。人类活动和1998年金沙江－长江特大洪水也促进了紫茎泽兰在该地区的扩散。另一方面，紫茎泽兰在新近入侵地区的垂直分布范围说明随海拔高度上升所引起的垂直地带性气候变冷是紫茎泽兰入侵和扩散的自然限制条件。 　　此外，通过对其余13种外来入侵植物入侵历史过程重建发现，除了牛膝菊、反枝苋和皱果苋最早入侵地位于内陆地区以外，其余10种外来入侵植物的最早入侵地都位于我国的东部、南部沿海和西南边境地区。这13种外来植物通常不是通过单一途径传入，而是通过两种或多种途径传入不同的地点。相应地，入侵后也呈现不同的扩散模式。这13种外来植物在我国大体上包括5种入侵和扩散式样即1）从南部沿海和西南边境分别向内陆扩散，如霍香蓟、飞机草，北美商陆;2）从东部沿海向内陆扩散，例如钻形紫菀，小蓬草、北美独行菜和喜旱莲子草;3）从东部沿海、南部沿海和西南边境分别向内陆扩散，如刺苋;4）从南部沿海向内陆扩散，如土荆芥;5）从最早入侵的内陆地区向周边地区扩散，如反枝苋、牛膝菊和皱果苋。分布区动态分析显示当前这13种外来入侵植物在我国都还处在扩散阶段，而且有些外来植物还处在快速扩散阶段，如飞机草。 　　在上述研究基础上，我们对这13种外来入侵植物开展了生态位模型适生区预测和比较分析。并且结合其扩散动态和当前分布现状，我们对其潜在的分布区及扩散趋势进行了系统分析和预测。飞机草、土荆芥、霍香蓟、牛膝菊和北美商陆，这5种外来植物的现有分布区明显小于其在我国的适生区，因此它们在我国的潜在分布区很广，应给予足够的重视。喜旱莲子草、反枝苋、皱果苋、北美独行菜、一年蓬、钻行紫菀和北美商陆在我国的入侵范围与其适生区域之间基本重合，但在其入侵范围内仍存在大面积的未入侵区域。这些未入侵地区通常被已入侵区域所包围，因此也应当给予足够的重视。刺苋和小蓬草在我国的潜在的分布区相对较小，这些植物已经基本上完全入侵了其适生区。因此，这些植物在我国向临近区域继续扩散的可能性不大。 　　

采用AFLP分子标记和分子生物学方法对资源植物的评价

中国资源植物丰富，蕴藏着优异的基因资源，开发和利用这些优异资源是植物学研究的重要课题。本文面向国家重大需求选择两种资源植物一羊草(Leymuschinensis (Trin.)Tzvel)和向日葵(llelia thus annuus L．)，采用分子标记技术和分子生物学方法对其进行评价和研究，以期为资源利用提供依据。由于两种植物本身的差别和采用的研究方法各异，故分别论述。 羊草，隶属禾本科赖草属，是欧亚大陆草原区东部重要建群种之一。羊草是牧草之王，是我国比较有优势的战略性生物资源，对我国北方畜牧业的发展以及生态环境的保育均具有重要意义。近年来，由于缺乏科学管理、过度放牧等不利影响，加之羊草本身固有的“三低”问题（即抽穗率低、结实率低、发芽率低）已对羊草生物多样性维持构成了严重的威胁，限制了我国人工草地建设和天然草地的改良及沙化治理的步伐。因此，如何通过形态调查结合生物技术手段评价羊草遗传多样性为建立核心种质及改良羊草、快速评价和创造新的种质、如何加快育种进程便成为当前亟待解决的问题。本文围绕这些问题开展了系统的研究并取得如下结果： 1． 对羊草的形态调查和AFLP分析，表明羊草是一种形态变异较大但是遗传变 异较小的物种。两种生态型的表现显著差异，其中灰绿生态型羊草比黄绿生 态型差异大。羊草遗传多样性与包括长期的栽培驯化、地理分布有很大的相 关性，地理来源相同的几乎全部聚到了一组。 2． 通过主成分分析和通径分析，简化了羊草31个性状分析的复杂性，了解到 羊草无性繁殖受好的营养生长促进。 3． AFLP分子标记技术在分析羊草遗传多样性方面有显著优势，尤其是对于羊 草这样多态性不高的物种是一种非常有效的分析工具。在分析AFLP数据时 采用聚类分析和主坐标分析相结合的方法，既兼顾了亲缘关系较近的种质之 间的关系调查也兼顾了亲缘关系较远的种质之间的关系调查。 4．羊草AFLP反应，不同引物所获得的总带数和多态性带数差别明显。羊草基因 组对3’端有选择性碱基TN的所有EcoRJ选择性引物扩增效果很差，前人 的有关赖草属的遗传研究也支持这一结果。 向日葵(n=17)，属于菊科( Compositae)向日葵属(Helia thus)，向日葵的研究重要领域是向日葵杂种生产，而细胞质雄性不育系的使用是杂种优势育种的核心。全世界90%以上的向日葵杂交种生产仍然在使用同一个细胞质类型PETI，玉米遗传单一给生产带来的毁灭性打击仍然令研究者和生产者记忆犹新，因此寻找更多的细胞质类型仍然是研究者的重要任务。本研究围绕一个新的不育源(G20023)的发现及鉴定，通过使用不育的G20023的保持系、恢复系、恢复的Fi代、回交一代之间比较以及与属于PETI细胞质类型的不育系的相应材料进行比较，找出与这一新的细胞质类型不育表型有关的可能差异序列，来探讨其不育机制，得到如下结果： 1、 通过田间杂交试验，证明G20023的保持系有很多（已证实有24份）， 目前找到的恢复系只有一个，H.maximiliani。G20023不育源作为一 种新的细胞质类型可以成为将来杂交育种的候选资源。同时，我们找 到一些表型证据，除了无花粉之外，G20023与PETI表型的典型不同 之处还在于前者的花药上下均为分离状态，而后者花药的基部联合， 顶部分离。显然，不同的细胞质类型在解剖结构上可能表现不尽相同。 2、 与线粒体基因组特异基因的核酸序列比较，结果表明，G20023线粒体 基因组上没有orfH522序列，与PETI表现出差异;此外，在基因atp6 位点也与PETI不同，而且在该位点也与同属向日葵ANTI不相同。同 时由于orf873并没有出现在ANTI中而出现在G20023中，因此我们可 以认为G20023这一个新的不育系是与ANTI和PETI不同的细胞质类 型。 3、 在参考常规线粒体DNA提取方法的基础上，我们做了很多改进，建立 了自己的向日葵线粒体DNA提取方法。该方法更快更简单，提取的线 粒体DNA完全可以用于酶切和杂交。 4、 G20023不育源由于其稳定的不育性状，可以作为培育无花粉彩色向日 葵杂交种的亲本材料，我们通过此不育源选育适当花色的无花粉观赏 向日葵生产杂交种。

不同放牧率对不同草原主要植物叶片解剖结构和化学成分的影响

本文选取不同放牧率下的优势植物为研究对象，主要从植物解剖结构和化学成分方面，对内蒙古典型草原和北美混合普列里草原的放牧演替机制进行探讨，这将有助于进一步揭示放牧演替过程及其主要植物对放牧的适应机理。具体研究结果如下： 1. 长期不同放牧率的放牧（内蒙古典型草原区放牧15年和混合普列里草原放牧19年）对所有物种（IMGERS的羊草（Leymus chinensis）、冰草（Agropyron cristatum）、糙隐子草（Cleistogenes squarrosa）、扁蓿豆（Melissitus rutenica）、小叶锦鸡儿（Caragana microphylla）、冷蒿（Artemisia frigida）和星毛委陵菜（Potentilla acaulis）和CGREC的Artemisia frgida ，Poa pratensis， Agropyron smithii，Solidago rigida， Helianthus rigidus和Symphoricarpos occidentalis）叶片的角质层厚度、表皮细胞面积、叶肉细胞面积、栅栏/海绵组织厚度、叶片厚度、中脉厚度均产生显著影响;放牧显著影响了两个研究区不同生活型功能群植物叶片的角质层厚度、表皮细胞面积、叶肉面积、栅栏/海绵组织和中脉厚度。内蒙古典型草原研究区草本植物功能群的叶片下角质层厚度、栅栏/海绵组织厚度显著大于灌木功能群，而美国混合普列里草原研究区，草本植物功能群的叶片下角质层厚度、表皮细胞面积、叶肉细胞面积、栅栏/海绵组织厚度、叶片厚度、中脉厚度均显著大于灌木功能群。 2. 内蒙古典型草原研究区放牧率显著影响了糙隐子草和小叶锦鸡儿的比叶面积（SLA）;在美国混合普列里草原研究区，放牧率显著影响了冷蒿SLA。但对生活型功能群的SLA影响不显著。 3. 内蒙古典型草原研究区放牧对物种叶片叶绿素含量、纤维素含量影响显著，放牧仅显著增加了扁蓿豆叶片的含氮量。美国混合普列里草原研究区物种叶片的全碳、干物质、酸性洗涤纤维、叶绿素a+b含量受放牧率的显著影响。两研究区放牧率对叶片叶绿素a+b含量影响显著。放牧率也显著影响了不同生活型功能群的全碳含量和叶绿素a+b含量。 4. 两研究区叶片表皮细胞面积和叶肉细胞面积无牧和重牧下显著正相关，表皮细胞面积和叶片厚度在轻牧下显著正相关，叶肉细胞面积和叶片厚度在无牧、中牧和重牧下显著正相关，比叶面积和叶绿素a+b含量在轻牧下显著正相关。扁蓿豆叶片的表皮细胞面积和叶肉细胞面积间存在显著的正相关，其全碳含量和叶绿素a+b含量间存在显著的正相关;Artemisia frgida叶片的角质层厚度和叶片厚度间显著正相关，Poa pratensis叶片的角质层厚度和表皮细胞面积间显著负相关，Solidago rigida叶片表皮细胞面积和光合速率显著正相关。Helianthus rigidus叶片叶肉细胞面积和叶片厚度显著负相关。

矿山环境中锑的表生地球化学研究—以贵州半坡锑矿区为例

锑(antimony，Sb)是一种典型的毒害重金属元素。我国作为世界上最主要的锑生产国，有关锑的表生地球化学及锑污染防治研究还很薄弱。贵州省位于我国西南低温成矿域中心，具有高Sb地球化学背景值，是我国重要的锑工业基地，境内分布着大量Sb矿床，锑矿储量居全国第四位。贵州又地处我国西南喀斯特中心区域，岩溶地貌极其发育，生态环境脆弱。由于大规模锑矿资源的开采利用，锑矿区土壤和水体中Sb污染十分突出，并通过水体或食物链对矿区及流域居民健康形成潜在威胁。因此，研究矿山环境中Sb的表生地球化学对于认识Sb污染的环境效应和寻求Sb污染修复方式具有至关重要的意义。 本文以贵州半坡大型锑矿区为研究对象，系统研究了Sb在矿区岩石、固体废弃物、土壤、水体和植物体等表生环境介质中的分布、迁移、富集规律与赋存机制，阐明了矿区Sb的表生地球化学过程规律，评价了矿区围岩、（废）矿石和尾矿砂等样品的产酸潜力，筛选出新的锑潜在超富集植物和超耐受性植物，并获得以下主要认识： （1） 独山半坡锑矿区是典型高锑地质地球化学背景区，不仅矿石中Sb含量很高，而且围岩中Sb含量也远高于上地壳Sb的平均含量。辉锑矿是锑的主要载体和释放源。 （2） 在固体废弃物中，冶炼废渣中Sb含量最高，其次为废石堆和尾砂库。废石中元素特征基本与围岩一致，继承了围岩的元素特征，而尾矿砂由于破碎、浮选过程的改造，其元素含量特征与围岩存在差异。尾砂砂中Sb以中层最富，底层和表层依次降低，与Fe、Al矿物的分布特征相似，这种分布模式主要受氧化还原条件、雨水淋滤、Fe/Al矿物吸附机制等因素控制。尾矿砂中不同相态的锑含量依次为残渣态>碳酸盐结合态>有机物结合态>可交换态>铁锰氧化物结合态，其中生物易利用态Sb的空间分布特征与总Sb分布特征一致。 （3） 所有的（废）矿石样品都具有潜在产酸能力，需要采取必要的措施抑制酸性矿山排水的产生。尾矿砂没有产酸潜力，这与矿石在选冶过程中加入石灰处理和S含量过低等原因有关，这种碱性环境可能有利于锑的表生地球化学活化和迁移。 （4） 研究区土壤受到严重锑污染，土壤中Sb含量高达51~7369 mg/kg，且随深度增加而逐渐降低，底层含量与对照区相近。这种高含量的Sb可能是与锑矿化有关的裸露岩石和土壤的自然风化淋滤过程和采矿、冶炼活动等造成的。研究区土壤中不同相态的Sb含量依次为残渣态>铁/锰氧化物结合态>碳酸盐结合态>有机物/硫化物结合态>可交换态。土壤中Sb的生物有效性很低。土壤中Sb的迁移与As密切相关。 （5） 研究区水体为SO4/HCO3-Ca型，水体中的Sb以溶解态、悬浮态和沉积物形式存在，并通过吸附-解吸附和水流推移方式进行迁移。洪水期和枯水期水体中Sb的扩散迁移机制存在差异，洪水期研究区内水体对岔河下游水体污染危害更大。在枯水期，岔河水体中溶解态Sb含量和沉积物中Sb含量存在显著正相关关系，表明枯水期沉积物和水体界面间达到物质交换平衡。水体的氧化还原条件和pH值控制着水体中Sb的存在形态。沉积物中不同相态Sb含量依次为残渣态>碳酸盐结合态，铁锰氧化物结合态>可交换态，有机物/硫化物结合态。研究区水体中Sb主要受矿区采矿、选冶活动的强烈影响。水体中硫同位素示踪研究发现，岔河下游水体中59%的S来自于矿山硫源的贡献，表明矿山活动严重影响到岔河下游水体水质。 （6） 植物对土壤中Sb的吸收与植物部位和种类有关。大体上地下部分>地上部分，且根>叶片>茎。在农作物中，白菜>甘蓝>辣椒和稻谷。研究发现，土荆芥(Chenopodium ambrosioides Linn.)是新的Sb潜在超富集植物，鬼针草(Bidens pilosa Linn.)、一年蓬(Erigeron annuus (Linn.) Pers.)、佛甲草(Sedum lineare Thun)、凹叶景天(Sedum emarginatum Migo)、灰灰菜(Chenopodium album Linn.)和鼠麹草(Gnaphalium affine D. Don)也能大量富集Sb，并有很强耐受性。 （7） 研究区这种特殊的高Sb地质环境和人为活动的影响，导致表生环境中Sb的高度富集，造成农作物、水体中Sb含量极高，通过食物链和水体严重影响当地居民的身体健康。

Biological wastes as plant growth substrate amendments

Globally, agriculture is being intensified with mechanization and increased use of synthetic fertilizers and pesticides. There has been a scaling up of production to satisfy the demands of supermarket distribution. Problems associated with intensification of production, trade globalisation and a larger market demand for greater volumes of fresh produce, include consumers' concern about pesticide residues and leaching of nutrients and pesticides into the environment, as well as increases in the transmission of human food-poisoning pathogens on raw vegetables and in fruit juices. The first part of this research was concerned with the evaluation of a biological control strategy for soil-borne pathogens, these are difficult to eliminate and the chemicals of which the most effective fumigants e.g. methyl bromide, are being withdrawn form use. Chitin-containing crustaceans shellfish waste was investigated as a selective growth substrate amendment in the field, in glasshouse and in storage trials against Sclerotinia disease of Helianthus tuberosus, Phytophthora fragariae disease of Fragaria vesca and Fusarium disease of Dianthus. Results showed that addition to shellfish waste stimulated substrate microbial populations and lytic activity and induced plant defense proteins, namely chitinases and cellulases. Protective effects were seen in all crop models but the results indicate that further trials are required to confirm long-term efficacy. The second part of the research investigated the persistence of enteric bacteria in raw salad vegetables using model food poisoning isolates. In clinical investigations plants are sampled for bacterial contamination but no attempt is made to differentiate between epiphytes and endophytes. Results here indicate that the mode isolates persist endophytically thereby escaping conventional chlorine washes and they may also induce host defenses, which results in their suppression and in negative results in conventional plate count screening. Finally a discussion of criteria that should be considered for a HACCP plan for safe raw salad vegetable production is presented.

Enhanced phytoextraction: I. Effect of EDTA and citric acid on heavy metal mobility in a calcareous soil

Phytoextraction, the use of plants to extract heavy metals from contaminated soils, could be an interesting alternative to conventional remediation technologies. However, calcareous soils with relatively high total metal contents are difficult to phytoremediate due to low soluble metal concentrations. Soil amendments such as ethylene diaminetetraacetate (EDTA) have been suggested to increase heavy metal bioavailability and uptake in aboveground plant parts. Strong persistence of EDTA and risks of leaching of potentially toxic metals and essential nutrients have led to research on easily biodegradable soilamendments such as citric acid. In our research, EDTA is regarded as a scientific benchmark with which degradable alternatives are compared for enhanced phytoextraction purposes. The effects of increasing doses of EDTA (0.1, 1, 10 mmol kg(-1) dry soil) and citric acid (0.01, 0.05,0.25,0.442, 0.5 mol kg(-1) dry soil) on bioavailable fractions of Cu, Zn, Cd, and Pb were assessed in one part of our study and results are presented in this article. The evolution of labile soil fractions of heavy metals over time was evaluated using water paste saturation extraction (similar to soluble fraction), extraction with 1 M NH4OAc at pH 7 (similar to exchangeable fraction), and extraction with 0.5 M NH4OAc + 0.5 M HOAc + 0.02 M EDTA atpH 4.65 (similar to potentially bioavailable fraction). Both citric acid and EDTA produced a rapid initial increase in labile heavy metal fractions. Metal mobilization remained constant in time for soils treated with EDTA, but metal fractions was noted for soils treated with citric acid. The half life of heavy metal mobilization by citric acid varied between 1.5 and 5.7 d. In the following article, the effect of heavy metal mobilization on uptake by Helianthus annutis will be presented.

Sexual System, Sex Ratio, and Group Living in the Shrimp Thor amboinensis (De Man): Relevance to Resource-Monopolization and Sex-Allocation Theories

The sexual system of the symbiotic shrimp Thor amboinensis is described, along with observations on sex ratio and host-use pattern of different populations. We used a comprehensive approach to elucidate the previously unknown sexual system of this shrimp. Dissections, scanning electron microscopy, size-frequency distribution analysis, and laboratory observations demonstrated that T amboinensis is a protandric hermaphrodite: shrimp first mature as males and change into females later in life. Thor amboinensis inhabited the large and structurally heterogeneous sea anemone Stichoclactyla helianthus in large groups (up to 11 individuals) more frequently than expected by chance alone. Groups exhibited no particularly complex social structure and showed male-biased sex ratios more frequently than expected by chance alone. The adult sex ratio was male-biased in the four separate populations studied, one of them being thousands of kilometers apart from the others. This study supports predictions central to theories of resource monopolization and sex allocation. Dissections demonstrated that unusually large males were parasitized by an undescribed species of isopod (family Entoniscidae). Infestation rates were similarly low in both sexes (approximate to 11%-12%). The available information suggests that T. amboinensis uses pure search promiscuity as a mating system. This hypothesis needs to be formally tested with mating behavior observations and field measurements on the movement pattern of both sexes of the species. Further detailed studies on the lifestyle and sexual system of all the species within this genus and the development of a molecular phylogeny are necessary to elucidate the evolutionary history of gender expression in the genus Thor.

Cys mutants in functional regions of Sticholysin I clarify the participation of these residues in pore formation

Experimental evidence shows that the mechanism of pore formation by actinoporins is a multistep process, involving binding of the water-soluble monomer to the membrane and subsequent oligomerization on the membrane surface, leading to the formation of a functional pore. However, as for other eukaryotic pore-forming toxins, the molecular details of the mechanism of membrane insertion and oligomerization are not clear. In order to obtain further insight with regard to the structure-function relationship in sticholysins, we designed and produced three cysteine mutants of recombinant sticholysin I (rStI) in relevant functional regions for membrane interaction: StI E2C and StI F15C (in the N-terminal region) and StI R52C (in the membrane binding site). The conformational characterization derived from fluorescence and CD spectroscopic studies of StI E2C, StI F15C and StI R52C suggests that replacement of these residues by Cys in rStI did not noticeably change the conformation of the protein. The substitution by Cys of Arg(52) in the phosphocholine-binding site, provoked noticeable changes in rStI permeabilizing activity; however, the substitutions in the N-terminal region (Glu(2), Phe(15)) did not modify the toxin`s permeabilizing ability. The presence of a dimerized population stabilized by a disulfide bond in the StI E2C mutant showed higher pore-forming activity than when the protein is in the monomeric state, suggesting that sticholysins pre-ensembled at the N-terminal region could facilitate pore formation. (C) 2011 Elsevier Ltd. All rights reserved.

Atributos bioquímicos e químicos do solo rizosférico e não rizosférico de culturas em rotação no sistema de semeadura direta

Sistemas autossustentáveis favorecem as populações microbianas devido à conservação e ao aumento da matéria orgânica no solo. Além disso, as plantas que fazem parte desses sistemas promovem o efeito rizosférico, por meio da zona de influência das raízes, que resulta no aumento da atividade e na modificação da população microbiana. O objetivo deste trabalho foi avaliar o efeito da rotação de culturas de inverno sobre sequências de verão, em sistema de semeadura direta, nos atributos bioquímicos (amilase, urease, celulase e protease) e químicos (carbono orgânico total - COT, carboidratos totais e proteínas totais) em solo rizosférico (SR) e não rizosférico (SNR). Este estudo foi constituído de três culturas de inverno: milho (Zea mays L.), girassol (Helianthus anuus L.) e guandu (Cajanus cajan (L.) Millsp), que estavam em rotação sobre três sequências de verão: soja/soja (Glycine max L.), milho/milho e soja/milho, e duas posições no solo: solo aderido às raízes das plantas (SR) e solo da entrelinha de plantio (SNR). As atividades da amilase, celulase, protease e urease no SR foram 16, 85, 62 e 100 % maiores do que no SNR; para COT e proteínas totais a diferença foi de 21 %. Das culturas de inverno, o milho foi a que mais estimulou as atividades das enzimas amilase, celulase, urease e protease no SR, bem como a atividade das enzimas amilase, urease e protease no SNR. de modo geral, os teores de proteínas totais não foram influenciados pelas culturas de inverno e pelas sequências de verão; os carboidratos totais foram influenciados pelas culturas de inverno milho e girassol. Para o COT houve influência apenas da sequência de verão milho/milho. Os atributos bioquímicos e químicos avaliados neste estudo podem ser utilizados como indicadores das alterações no solo promovidas pelas culturas de inverno e pelas sequências de verão.

Desenvolvimento de grãos e produção de fitomassa do girassol em função de adubações boratadas

Sunflower crop is a good option to no-tillage system; however, it is sensible to low B availability, what is common in tropical soils. The objective of this study was to evaluate the crop development, grain yield and grain size as affected by B fertilizer doses applied in the sowing and by leaf application. The experiment was conducted in controlled conditions, with four Borax doses (0; 1; 2 e 3 kg B ha(-1)) in the sowing interacting with four Boric Acid doses (0; 0,5; 1 e 2 kg B ha(-1)) by leaf application. The plants were managed until grain physiological maturity, when stems, leaves and capitula were sampled, besides the counting and weighing of two grain classes separated by size. The B fertilizer applied in the sowing lines increased total dry matter phytomass and grain yield. However, when interacting with high doses of B by leaf application, these values decreased. Grain size was increased with leaf fertilization, when B was not applied in the soil or with only 1 kg B ha(-1) in the sowing line. on the other hand, when B was applied from 2 to 3 kg ha(-1) in the crop sowing, high doses of leaf fertilizer affected grain filling.

Avaliação de híbridos de girassol e relação entre parâmetros produtivos e qualitativos

O presente estudo teve o objetivo de determinar genótipos superiores de girassol, bem como realizar estudos de correlação entre suas características. Avaliou-se seis híbridos de girassol em condições de safrinha no município de Palotina - PR. O estudo avaliou quinze variáveis: massa seca de folhas, massa seca de caule e pecíolos, massa seca total, área foliar, altura de planta, diâmetro de colmo, massa de grãos por capítulo, diâmetro de capítulo, percentagem de grãos normais, massa de mil grãos, número de grãos por capítulo, produtividade, teor de proteína bruta, teor de óleo e rendimento de óleo. Os dados direcionam os híbridos H360 e MG2 com boa produtividade e maior teor de óleo para a produção de óleo e os híbridos M734 e Aguará 3 para a alimentação animal, com boa produtividade e menor teor de óleo. As correlações entre produtividade e os componentes de produção foram de 0,62; 0,47; 0,60; 0,49 e 0,47 para massa de grãos por capítulo, diâmetro de capítulo, percentagem de grãos normais, massa de mil grãos e número de grãos por capítulo, respectivamente, concluindo que a seleção de materiais a partir desses componentes ocasionará a seleção de materiais promissores em produtividade.

Estabilidade de agregados do solo após manejo com rotações de culturas e escarificação

O objetivo desse trabalho foi avaliar, em solo compactado, a estabilidade dos agregados influenciada pelo cultivo de espécies de cobertura em esquema de rotações de culturas, em sistema de semeadura direta, mediante o efeito da escarificação mecânica. As rotações de culturas repetidas por três anos consecutivos envolveram o cultivo de triticale e girassol, no outono-inverno, associados ao cultivo de milheto, de sorgo forrageiro e de Crotalária júncea como plantas de cobertura, antecedendo o da soja (cultura de verão). No tratamento envolvendo a escarificação mecânica, a área permaneceu em pousio entre os cultivos de outono-inverno e de verão. O experimento foi realizado na Fazenda Experimental Lageado (Botucatu-SP), nos anos agrícolas de 2003/2004, 2004/2005 e 2005/2006. O delineamento experimental foi o de blocos ao acaso, com quatro repetições, em esquema de parcelas subdivididas. As amostras para a análise da estabilidade de agregados foram coletadas nas profundidades de 0 a 0,05 m e de 0,05 a 0,10 m após o manejo das plantas de cobertura em dezembro de 2003 e de 2005. Logo no primeiro ano de instalação do experimento, o cultivo de triticale resultou em maior porcentagem de agregados com mais de 2 mm, maior DMG e maior DMP na camada de 0 a 5 cm, além de maior DMP na camada de 0,05 a 0,10 m. Já a escarificação do solo e a ausência do cultivo de plantas de cobertura proporcionaram menor porcentagem de agregados maiores que 2 mm e menor DMP na camada de 0,05 a 0,10 m. A estabilidade dos agregados foi influenciada pela rotação de culturas, sendo maior na camada de 0 a 0,05 m e de 0,05 a 0,10 m quando o triticale foi introduzido como espécie de outono-inverno.

Production and action pattern of inulinase from Aspergillus Niger-245: hydrolysis of inulin from several sources

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)