Eficácia do glyphosate e 2,4-d no controle de commelina villosa

The aim of this study was to evaluate the efficacy of glyphosate and 2,4-D alone and in combination, in the control of Commelina villosa. We studied two methodologies for evaluating herbicide absorption in eight time intervals for washing (simulating rainfall after application) and cutting of leaves (simulating abortion as a defense strategy): 2, 4, 6, 8, 12, 24 and 48 hours after herbicide application, and a treatment without washing or cutting the leaves in a completely randomized design with four replications in a 3 x 7 + 1 factorial design (three herbicides x seven periods – hours after application). Herbicides and doses tested were: glyphosate (1,440 g ha-1), 2,4-D (720 g ha-1) and a mixture of glyphosate + 2,4-D (1,080 + 720 g ha-1). The simulation of rain interfered negatively in the plant control with glyphosate. The control with the herbicide 2,4-D was affected only for the period of 2 hours. Periods of rain simulation did not influence the control of plants with a mixture of glyphosate + 2,4-D. For the study with the cutting of treated leaves, all treatments regardless of the period of cutting the leaves were influenced negatively in terms of plant control, the plants showing regrowth when treated with 2,4-D alone.

Calamagrostis arundinacea x villosa n. hybr. : C. indagata Torges & Hskn.

Dr. Torges

Ultramorphology and histochemistry of fat body cells from last Instar larval of the Pachycondyla (=Neoponera) villosa (Fabricius) (Formicidae: Ponerinae)

As células do corpo gorduroso de Pachycondyla (=Neoponera) villosa distribuem-se como uma única camada entre a cutícula e o trato digestivo, formando um conjunto de células agrupadas e recobertas por uma fina membrana. Não foram identificados tipos celulares distintos por meio da ultramorfologia, porém a histologia revelou três tipos celulares distintos: os trofócitos, mais abundantes, as células de urato, distribuídas por entre os trofócitos, e os enócitos, menos abundantes que os demais. Os enócitos são comumente observados próximos da cutícula. Histoquimicamente, os trofócitos apresentaram reação positiva para proteínas básicas no núcleo e no citoplasma e reação fortemente positiva nos grânulos citoplasmáticos. O teste para carboidratos foi fortemente positivo em todo o citoplasma, enquanto para os lipídeos observou-se reação positiva nas vesículas citoplasmáticas. em relação às células de urato, estas apresentaram reação positiva para proteínas básicas no núcleo e citoplasma, por entre as vesículas. Essas células não apresentaram reação para o teste de PAS e Sudan Black B. Quanto aos enócitos, estes apresentaram citoplasma fracamente positivo ao PAS e fortemente positivo ao Sudan Black B e para o azul de bromofenol.

An ethnobotanical study of traditional anti-inflammatory plants used by the Lohit community of Arunachal Pradesh, India

Aim of the study: Most people especially in rural areas depend on herbal medicines to treat many diseases including inflammation-related ailments such as rheumatism, muscle swelling, cut wound, accidental bone fracture, insect bites, pains and burn by fire and hot water. The objectives of this study were: to catalog ethno-medicinal plants of Lohit community, ecological status, indigenous folk medicinal uses, morphological parts used and to determine their reported pharmacological studies. Materials and methods: The ethnobotanical information on traditional medicinal plants exclusively used for management of inflammation-related ailments by the Khampti community of Arunachal Pradesh, India was based on first-hand field survey work through semi-structured interviews. Results and conclusion: A total of 34 species in 32 genera and 22 families were encountered during the field survey. Botanical families such as Asteraceae, Euphorbiaceae, Zingiberaceae and Lamiaceae were represented by the highest numbers of species reported in this study. Thirteen plant species, namely: Bombax ceiba, Canarium strictum, Chloranthus erectus, Xanthium indicum, Lycopodium clavatum, Coleus blumei, Batrachospermum atrum, Chlorella vulgaris, Marchantia palmata, Marchantia polymorpha, Eria pannea, Sterculia villosa and Alpinia galanga are reported for the first time for the treatment of inflammation-related diseases.

An inventory of the ethnobotanicals used as anti-diabetic by a rural community of Dhemaji district of Assam, Northeast India

Ethnopharmacological relevance: Traditional remedies used for treating diabetic ailments are very important in the primary health care of the people living in rural Dhemaji district of Assam, north-east India. Novel information gathered from the current survey is important in preserving folk indigenous knowledge. Materials and methods: Interviews were conducted amongst 80 households comprising of 240 individuals using semi-structured questionnaires. The focus was on plants used in treating diabetes mellitus. Results: The current survey documented 21 plant species (20 families) which are reportedly used to treat diabetes mellitus by the rural people in the study area. To the best of our knowledge, Amomum linguiforme, Cinnamomum impressinervium, Colocasia esculenta, Dillenia indica, Euphorbia ligularia, Garcinia pedunculata, Solanum indicum, Sterculia villosa and Tabernaemontana divaricata are recorded for the first time based on globally published literature as medicinal plants used for treating diabetes mellitus and related symptoms. Conclusions: The wide variety of plants that are used to treat diabetes mellitus in this area supports the traditional value that medicinal plants have in the primary health care system of the rural people of Dhemaji district of Assam. The finding of new plant uses in the current study reveals the importance of the documentation of such ethnobotanical knowledge. (C) 2011 Elsevier Ireland Ltd. All rights reserved.

Recent freshwater Ostracoda and Bivalvia from Indo-Nepal border (Tethys Himalaya): Ecological implications

We report for the first time Recent ostracods and bivalves from Central Tethys Himalaya, collected from Changru village, which is situated in the Tinker valley of northwestern Nepal near the Indo-Nepal border. The specimens were collected from the surface sediments of a small pond, shaped by a small tributary of the glacier fed Tinker River. The ostracod species belonging to the families Ilyocyprididae, Cyprididae and Candonidae and one bivalve (Pisidium sp., Family Pisidiidae) have been identified. Psychrodromus olivaccus and Potamocypris villosa are being reported for the first time from India and Nepal. Several broken and unidentifiable gyrogonites of charophyta were also recovered. The ostracod sample, as a whole, points to a shallow freshwater lake environment influenced by slowly running carbonate rich waters under cool temperatures, low mineralization and sparse vegetation. This is in accordance with the occurrence of Pisidium, which is commonly associated with ostracods in the freshwater lakes and streams. The ostracod fauna shows affinity with the fossil and extant forms recovered from the Higher and Tethys Himalaya of NW India. This opens a new opportunity for studying ostracods in the Indian Central Himalaya - a region which otherwise has been ignored until now.

丁香属（Syringa L.）的分类学修订

丁香属隶属于木犀科，分布于东南欧和东亚至喜马拉雅地区，我国是丁香属的现代分布中心。《中国植物志》（61卷，1992）记录了我国野生丁香种类16种;《Flora of China》（15卷，1996）记录了中国原产丁香种类16种，并认为全世界大约有20种。丁香属属下分类等级划分分歧较大，很多种的划分也存在争议。花叶丁香、四川丁香等种类是根据栽培植物描述的，没有指定模式标本，给分类处理带来了一定困难。另外，丁香属很多分类群的性状变异非常复杂，仅根据有限的标本很难做出合理的分类处理。本研究通过广泛查阅文献和标本，同时进行野外居群取样和性状观察，对各类群的性状在居群内和居群间的变异进行统计学分析，判断其分类价值，并运用多变量分析的方法，为各类群的合理划分提供依据。结合性状分析和地理分布等证据，做出分类处理。 作者查阅了国内外16个标本馆的近2000份标本，其中模式标本约70份。对我国12个省市的40余个居群进行了取样和观察，采集标本500余份，涉及了《中国植物志》61卷收录的除了藏南丁香以外的所有类群。通过对9个复合体的40余个性状在居群内和居群间的变异进行统计分析，发现叶片类型、叶柄长度、花序着生类型、花冠大小、花丝长度、花药颜色在不同类群间差异明显，可以用作划分种的依据;叶片形状、叶片毛被、花序轴毛被、花冠管形状等性状在有些复合体内的居群间呈现连续的变异，只能用作种下等级（亚种）的划分;叶片大小、花序轴形状、花药着生在花冠的位置、蒴果是否被皮孔等性状在不同复合体的居群间呈现间断或连续的变异，视不同情况可以用作种间或种下等级的划分依据，或作种内变异处理;而叶脉、花色、花萼齿裂、花冠裂片形状等性状在居群间差异不大，不适合用作分类依据。 在性状分析和多变量分析的基础上，本文将丁香属划分为2组2系12种13亚种，其中短花冠管组有1种3亚种;长花冠管组的顶生花序系有5种5亚种，侧生花序系有6种5亚种，并指定了各组和系的模式种;编制了属下各组、系、种和亚种划分的检索表，对12种13亚种进行了形态描述、标本引证，给出了地理分布图和生境，并提出了分类处理依据。文中对巧玲花、皱叶丁香、红丁香和云南丁香等复合体内的一些分类群进行了归并，做出4个新组合：S. pubescens ‘Meyer’、S. villosa subsp. wolfii、S. yunnanensis subsp. sweginzowii和S. yunnanensis subsp. tomentella，处理了11个新异名（S. fauriei H. Lév.、S. julianae C. K. Schneid.、S. meyeri var. spontanea M. C. Chang、S. pinetorum W. W. Sm.、S. wardii W. W. Sm.、S. oblata var. donaldii R. B. Clark et J. L. Fiala、S. afghanica C. K. Schneid.、S. protolaciniata P. S. Green et M. C. Chang、S. tibetica P. Y. Bai、S. reflexa C. K. Schneid. 、S. wilsonii C. K. Schneid.）。作者指定了3种4亚种(S. reticulata subsp. reticulata、 S. reticulata subsp. amurensis、S. pubescens subsp. microphylla、S. oblata subsp. dilatata的后选模式，并对其它10个名称指定了后选模式。文中还提出了分类处理原则，对我国丁 香属的分布及各地方植物志的记载进行了评述，并对丁香属的分布格局提出了作者的看法。 作者还对13个分类群的16号材料进行了染色体观察，发现除了毛丁香有染色体2n=48外，其它均为2n=46，其中朝阳丁香的染色体数目为首次报道。对小叶巧玲花不同异名（包括小叶蓝丁香、小叶巧玲花与小叶蓝丁香的杂交种）的材料进行染色体观察时，发现它们之间差异很小，进一步佐证了作者将其合并的合理性。另外野生的花叶丁香（华丁香）与栽培的花叶丁香在染色体数目上也无差异，支持了作者认为二者为同物异名的观点。

山黑豆属(豆科)的分类学修订

山黑豆属Dumasia DC.隶属于豆科、菜豆族、大豆亚族，广泛分布于亚洲、非洲东南部的热带、亚热带地区以及大洋洲的巴布亚新几内亚地区，其中大多数种在我国均有分布。该属形态变异复杂，命名混乱，缺乏世界性的系统分类学研究基础。基于这样的背景，本论文通过开展形态学、解剖学、细胞学及孢粉学等方面的综合研究工作，对该属进行了全面的分类修订，对属内种间关系及地理分布格局进行了分析和探讨，结果如下： 1、形态学 在研究了大量腊叶标本和进行了野外考察的基础上，对山黑豆属植物的形态性状的变异式样进行了分析。该属植物的根系形态较为多样，可以作为划分种的辅助形状，分为有主根类型 (2个种，心叶山黑豆D. cordifolia Benth. ex Baker和柔毛山黑豆D. villosa DC.) 和无主根类型 (5个种，云南山黑豆D. yunnanensis Y. T. Wei & S. K. Lee、小鸡藤D. forrestii Diels、硬毛山黑豆D. hirsuta Craib、山黑豆D. truncata Siebold & Zucc.和长圆叶山黑豆D. henryi (Hemsl.) B. Pan & X. Y. Zhu)；该属几乎全为多年生缠绕草本，只有柔毛山黑豆为半灌木，因此可能是该属中较原始的种类；该属植物的萼筒管状、筒口斜截形及花柱细长且弯曲处膨大等独特的解剖结构是区别该属其它近缘属的重要性状；花序和复叶的长度及荚果内种子数目变异幅度极大，缺乏稳定性，并非好的分类性状。 2、解剖学 应用光学显微镜，首次对山黑豆属7种植物 (心叶山黑豆、柔毛山黑豆、云南山黑豆、小鸡藤、硬毛山黑豆、山黑豆和长圆叶山黑豆) 的叶表皮进行了观察。结果表明，该属植物叶表皮细胞均呈不规则形，垂周壁为波状，气孔器类型以平列型为主。心叶山黑豆、柔毛山黑豆和云南山黑豆上表皮细胞垂周壁呈深波状；小鸡藤、长圆叶山黑豆、山黑豆和硬毛山黑豆上表皮细胞垂周壁呈浅波状。在一定程度上，上表皮细胞垂周壁的形态反映了山黑豆属种间的近缘关系。 3、细胞学 首次报道了山黑豆属3个种 (云南山黑豆、小鸡藤和硬毛山黑豆) 的染 色体数目，它们均为2n=20。在已报道染色体的植物种类中 (共计6种)，仅心叶山黑豆染色体数目为2n=22，其余5种 (柔毛山黑豆、山黑豆、云南山黑豆、小鸡藤和硬毛山黑豆) 染色体数目均为2n=20。与大豆亚族大多数属染色体数目为2n=22相比较，心叶山黑豆可能是该属中较为原始的种。 4、孢粉学 在光学显微镜下，对该属7种植物 (心叶山黑豆、柔毛山黑豆、云南山黑豆、小鸡藤、硬毛山黑豆、山黑豆和长圆叶山黑豆) 的花粉进行了观察，结果表明：该属植物为三角形6孔花粉，极轴短，每个角的萌发孔由一对小孔组成；外壁具网纹，网脊粗，网眼不规则。柔毛山黑豆和小鸡藤极面的网脊几乎不断裂；心叶山黑豆、云南山黑豆、硬毛山黑豆、山黑豆和长圆叶山黑豆极面的网脊断裂较多。外壁的纹饰在一定程度上反映了该属植物的近缘关系。 5、地理分布格局 该属植物主要分布在中国南部和西南，只有柔毛山黑豆分布到非洲和大洋洲。研究结果表明，具主根的2个种 (心叶山黑豆和柔毛山黑豆) 主要分布在中国西南部，其分布区域相互重叠；而无主根的5个种 (云南山黑豆、小鸡藤、硬毛山黑豆、山黑豆和长圆叶山黑豆) 由中国西南部向东到日本、韩国都有分布，分布区域几乎不重叠。本研究认为：有主根的心叶山黑豆和柔毛山黑豆可能代表了该属的原始类群；有主根类群衍生出其余无主根类型的各种，并由中国西南扩散到其他地区。 综合以上资料，本研究对山黑豆属进行了分类修订，认为该属含8种、1亚种及2变种，其中建立新组合1个，归并6个类群，并指定了4个后选模式。另外，本研究同时提供了该属植物的检索表、种的形态描述和插图以及地理分布图等信息。

联合固氮菌Klebsiella oxytoca SA2和水稻幼苗的相互作用

从内蒙流沙地先锋植物沙竹（Psammochloa mongokca）内分离到一株内生细菌，经鉴定定名为Klebsiella oxytoca SA-2 K.oxytoca SA-2兼性厌氧固氮，NH4+抑制其固氮酶合成。部分抑制固氮酶活性;N03 -抑制其固氮酶的合成和活性。 60℃灭活K.oxytoca SA-2整体菌免疫兔子得到抗血清。免疫印迹表明此抗血清具K.oxytoca SA-2种特异性。石蜡切片免疫金银染色结合显微观察发现K.oxytoca SA-2定殖于沙竹叶鞘薄壁细胞和叶片的薄壁细胞内。 K.oxytoca SA-2在半固体培养基中接种水稻幼苗，限菌培养21天，根内重新分离的数量达l06 cfu/g．FWroot，但K.oxytoca SA-2在富养的土壤中生长良好，表现为兼性内生菌。 限菌培养水稻（Oryza sativa）幼苗，石蜡切片免疫金银染色结合显微观察研究了的K.oxytoca SA-2的侵染特性．K.oxytoca SA-2可以通过侧根发生处和表皮细胞胞间层进入根内，在皮层薄壁细胞间隙大量定殖，在解体和看似完整的薄壁细胞内也有定殖，在根和茎中柱内K.oxytoca SA-2进入了木质部导管。在根基，K.oxytoca SA-2大量侵入了已解体的内皮层和中柱鞘细胞，植物细胞在K.oxytoca SA-2侵入后解体，可能表现为严格的局部超敏反应。 接种K.oxytoca SA-2 21天，水稻地上苗部分没有发现肉眼和显微可见的病症。与对照相比，接种K.oxytoca SA-2显著促进限氮培养水稻幼苗的生长。由于K.oxytoca SA-2在限碳限氮培养基和水稻幼苗共培养时能分泌NH4+和植物激素，它可能通过向水稻幼苗提供氮素和分泌植物激素促进植物生长。而且用固氮酶铁蛋白抗血清进行免疫金银染色发现定殖在根基皮层薄壁细胞胞间层和细胞间隙，木质部导管和茎基木质部导管的K.oxytoca SA-2可以表达固氮酶，固氮参与了K.oxytoca SA-2在水稻幼苗中的内生。 培养基内碳源（苹果酸）和培养温度对K.oxytoca SA-2和水稻幼苗相互作用的影响也进行了研究。 研究表明，K.oxytoca SA-2作为兼性内生固氮菌，能够和植物紧密联合，并在植物体内开拓一个有利的生态位固氮，而且K.oxytoca SA-2可以分泌NH4+和植物激素，在和植物相互作用中使植物受益。

小麦—偏凸山羊草6Mv/6B代换系的研究

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.

簇毛麦（ Dasypyrum villosum）矮秆突变体的研究

株高是农作物的重要农艺性状之一，适度矮化有利于农作物的耐肥、抗倒、高产等。20世纪50年代，以日本的赤小麦为矮源的半矮秆小麦的培育和推广，使得世界粮食产量显著增长，被誉为“绿色革命”。迄今为止，已报到的麦类矮秆、半矮秆基因已达70多个，但由于某些矮源极度矮化或者矮化的同时伴随不利的农艺性状，使得真正运用于育种实践的矮源较少。因此，发掘和鉴定新的控制麦类作物株高的基因，开展株高基因定位、克隆及作用机理等方面的研究，对实现麦类作物株高的定向改良，具有重要的理论意义和应用价值。簇毛麦(Dasypyrum villosum，2n＝14，VV)是禾本科簇毛麦属一年生二倍体异花授粉植物，为栽培小麦的近缘属。本课题组在不同来源的簇毛麦杂交后代中发现了一株自然突变产生的矮秆突变体。观察分析了该突变体的生物学特性，对矮秆性状进行了遗传分析，对茎节细胞长度、花粉的活力进行了细胞学观察，考察了该突变体内源赤霉素含量及不同浓度外施赤霉素对突变体的作用，分析了赤霉素生物合成途径中的内根贝壳杉烯氧化酶（KO）和赤霉素20氧化酶(GA20ox)的转录水平，对赤霉素20氧化酶和赤霉素3-β羟化酶(GA3ox)进行了克隆和序列分析，并对GA20ox进行了原核表达和表达的组织特异性研究。主要研究结果如下：1. 该突变体与对照植株在苗期无差异，在拔节后期才表现出植株矮小，相对对照植株，节间伸长明显受到抑制，叶鞘长度基本不变。在成熟期，对照植株的平均株高为110cm，而突变株的平均株高为32cm，仅为对照植株的1/3 左右。除了株高变矮以外，在成熟后期，突变株还表现一定程度的早衰和雄性不育。I2-KI染色法观察花粉活力结果表明，对照植株花粉90%以上都是有活力的，而突变植株的花粉仅20%左右有活力。2. 突变株与对照植株的杂交F1代均表现正常株高，表明该突变性状为隐性突变。F1代植株相互授粉得到的168株F2代植株中，株高出现分离，正常株高（株高高于80cm）与矮秆植株（株高矮于40cm）的株数比为130：38，经卡方检验，其分离比符合3：1的分离比，因此推测该突变体属于单基因的隐性突变。3. 用ELISA方法检测突变株和对照植株的幼嫩种子中内源性生物活性赤霉素（GA1＋3）含量，结果表明突变株的赤霉素含量为36 ng/ml，而对照植株的赤霉素含量为900 ng/ml。对突变株外施赤霉素，发现矮秆突变株的株高和花粉育性均可得到恢复。这些结果表明该突变株为赤霉素缺陷型突变。4. 用荧光定量PCR方法比较突变株与对照植株中内根贝壳杉烯氧化酶和赤霉素20氧化酶的转录水平，结果表明突变株的KO转录水平比对照植株分别提高了6倍（苗期）和16倍（成熟期），突变株的GA20ox转录水平与对照植株在苗期无明显差异，在成熟期突变株较对照植株则提高了10倍左右。这些结果表明该矮秆突变体与赤霉素的生物合成途径密切相关，而且极有可能在赤霉素的生物合成途径早期就发生了改变。5. 以簇毛麦总基因组为模板，同源克隆了GenBank登录号为EU142950，RT-PCR分离克隆了簇毛麦的GA3ox基因cDNA全长序列，分析结果表明该cDNA全长1206bp，含完整编码区1104bp，推测该序列编码蛋白含368个氨基酸残基，分子量为40.063KD，等电点为6.27。预测的氨基酸序列含有双加氧酶的活性结构，在酶活性中心2个Fe离子结合的氨基酸残基非常保守。该序列与小麦、大麦和水稻的GA3ox基因一致性分别为98%、96%、86%。基因组序列与cDNA序列在外显子部分一致，在478-715bp和879-1019bp处分别含238bp和140bp的内含子。6. 通过RT-PCR技术克隆了簇毛麦的GA20ox基因全长，命名为DvGA20ox，GenBank登录号为EU142949。该基因全长1080个碱基，编码359个氨基酸，具有典型的植物GA20ox基因结构。该基因编码的蛋白质与小麦、大麦、黑麦草等GA20ox蛋白的同源性分别为98%，97% 和91%。该序列重组到原核表达载体pET-32a(+)上，将获得的重组子pET-32a(+)-DvGA20ox转化大肠杆菌BL21pLysS后用IPTG进行诱导表达。SDS-PAGE分析表明，DvGA20ox基因在大肠杆菌中获得了高效表达，融合蛋白分子量为55kDa。定量PCR分析表明，该基因在簇毛麦不同器官中的表达差异明显：叶片中表达水平最高，根部表达水平次之，茎部和穗中表达较弱。在外施赤霉素后，该基因的表达水平在两小时以后急剧下降，表明该基因的表达受自身的反馈调节。本研究结果认为，(1)该簇毛麦矮秆突变体为单基因的隐性突变；(2)该矮秆突变体为赤霉素敏感突变，内源赤霉素含量检测表明突变体的内源性赤霉素含量仅为对照植株的1/30；(3)荧光定量PCR结果表明突变株的赤霉素生物合成途径的关键酶基因表达水平比对照植株高，而且突变植株的赤霉素生物合成改变很可能发生在赤霉素生物合成途径的早期；(4)GA20ox有表达的组织特异性，且受到自身产物的反馈调节。 Plant height is an impotrant agronomic trait of triticeae crops.Semi-dwarf cropcultivars, including those of wheat, maize and rice, have significantly increased grainproduction that has been known as “green revolution”. The new dwarf varieties couldraise the harvest Index at the expense of straw biomass, and, at the sametime, improvelodging resistance and responsiveness to nitrogen fertilizer. Moreover, dwarf traits ofplant are crucial for elucidating mechanisms for plant growth and development aswell. In many plant species, various dwarf mutants have been isolated and theirmodles of inheritance and physiology also have been widely investigated.The causesfor their dwarf phenotypes were found to be associated with plant hormones,especially, gibberellins GAs.Dasypyrum villosum Candargy (syn.Haynaldia villosa) is a cross-pollinating,diploid (2n = 2x = 14) annual species that belongs to the tribe Triticeae. It is native toSouthern Europe and West Asia, especially the Caucasuses, and grows underconditions unfavorable to most cultivated crops. The genome of D. villosum,designated V by Sears, is considered an important donor of genes to wheat for improving powdery mildew resistance, take-all, eyespot, and plant and seed storageprotein content. A spontaneous dwarf mutant was found in D. villosum populations.The biological character and modles of inheritance of this dwarf mutant are studied.The cell length of stem cell is observed. The influence of extraneous gibberellin tothe dwarf mutant is also examined; the transcript level of key enzyme of gibberellinbiosynthesis pathway in mutant and control plants is compared. GA3ox and GA20oxare cloned and its expression pattern is researched.1. The dwarf mutant showed no difference with control plants at seedlingstage.At mature stage, the average height of control plants were 110cm and the dwarfplants were 33cm. The height of the mutant plant was only one third of the normalplants due to the shortened internodes. Cytology observation showed that theelongation of stem epidermal and the parenchyma cells were reduced. The dwarfmutant also shows partly male sterile. Pollen viability test indicates that more than80% of the pollen of the mutant is not viable.2. The inheritance modle of this dwarf mutant is studied. All The F1 plantsshowed normal phenotype indicating that the dwarfism is controlled by recessivealleles. Among the 168 F2 plants, there are 130 normal plants and 30 dwarf plants, thesegregation proportion accord with Mendel’s 3:1 segregation. We therefore proposethat this dwarf phenotype is controlled by a single recessive gene.3. Quantitative analyses of endogenous GA1+3 in the young seeds indicated thatthe content of GA1+3 was 36ng/ml in mutant plants and 900ng/ml in normal plants.The endogenous bioactive GA1+3 in mutant plants are only about 1/30 of that innormal plants. In addition, exogenously supplied GA3 could considerably restore themutant plant to normal phenotype. These results showed that this mutant wasdefective in the GA biosynthesis.4. More than ten enzymes are involved in GA biosynthesis. KO catalyzes thefirst cytochrome P450-mediated step in the gibberellin biosynthetic pathway and themutant of KO lead to a gibberellin-responsive dwarf mutant. GA20ox catalyze therate-limited steps so that their transcript level will influence the endogenous GAbiosynthesis and modifies plant architecture. The relative expression levels of genesencoding KO and GA20ox were quantified by real time PCR to assess whether thechanges in GA content correlated with the expression of GA metabolism genes andwhere the mutant occurred during the GA biosynthesis pathway. In mutant plants,the transcript levels of KO increased about 6-fold and 16-fold at the seedling stage and elongating stage respectively comparing with the normal plants. For theseedlings, there was no notable difference in the expression of GA20ox betweenmutant and normal plants. At the elongating stage, GA20ox transcript increased 10times in mutant plants, suggesting that the GA biosynthesis pathway in mutant plantshad changed from the early steps rather than the late steps.5. A full length cDNA of D. villosum gibberellin 3β-hydroxylase homology(designated as DvGA3ox) was isolated and consisted of 1206bp containing an openreading frame of 1104bp encoding 368 predicted amino acid residues. Identityanalysis showed that the gibberellin 3β-hydroxylase nucleotide sequence shared 98%,96% and 86% homology with that of wheat, barley and rice. The predicted peptidecontained the active-site Fe of known gibberellin 3β-hydroxylase and the regionhomologous to wheat, barley and Arabidopsis. The genomic clone of gibberellin3β-hydroxylase has two introns.6. The full-length cDNA of D. villosum gibberellin 20 oxidase (designated asDvGA20ox) was isolated and consisted of 1080-bp and encoded 359 amino acidresidues with a calculated mol wt of 42.46 KD. Comparative and bio-informaticsanalyses revealed that DvGA20ox had close similarity with GA20ox from otherspecies and contained a conserved LPWKET and NYYPXCQKP regions. Tissueexpression pattern analysis revealed DvGA20ox expressed in all the tissues that wereexamined and the highest expression of DvGA20ox in expanding leaves followed byroots. Heterologous expression of this cDNA clone in Escherichia coli gave a fusionprotein that about 55KD. Transcript levels of DvGA20ox dramatically reduced twohours after application of biologically active GA3, suggesting that the biosynthesis ofthis enzymes might be under feedback control.

Datos sobre la vegetación de fuentes y arroyos de aguas nacientes en las montañas de la cornisa vasco-cantábrica

El presente trabajo recoge los estudios realizados por los autores en las comunidades vegetales que pueblan las fuentes y arroyos de los pisos colino y montano en la región vasco-cantábrica. La relación de citas bibliográficas existentes anteriormente y la contrastación con estudios similares en otros puntos de la zona y regiones próximas. Se ha prestado mayor interés a aquellas comunidades helofíticas caracterizadas por la presencia de Soldanella villosa Darracq. notable endemismo vasco-cantábrico.

Temporal and spatial appearance of the membrane cytoskeleton and perineuronal nets in the rat neocortex.

Parvalbumin-immunoreactive interneurons are surrounded by perineuronal nets, containing molecules of the extracellular matrix (e.g. tenascin-R). Furthermore, they seem to have a special cytoskeleton composed of, among others, ankyrinR and beta Rspectrin. In the present developmental study we showed that the intracellular markers parvalbumin, ankyrinR and beta Rspectrin as well as Vicia Villosa agglutinin, an extracellular marker for perineuronal nets, appeared in the second postnatal week. In the third postnatal week, ankyrinR and beta R spectrin were present in the parvalbumin-positive interneurons. Tenascin-R appeared in a similar topographic distribution as the intracellular markers. The adult pattern was established upon the end of the fourth postnatal week. Our results indicate that cytoskeletal maturity maybe a prerequisite for the organization of perineuronal nets of extracellular matrix.

Simple sequence repeats reveal uneven distribution of genetic diversity in chloroplast genomes of Brassica oleracea L. and (n=9) wild relatives

Diversity in the chloroplast genome of 171 accessions representing the Brassica 'C' (n = 9) genome, including domesticated and wild B. oleracea and nine inter-fertile related wild species, was investigated using six chloroplast SSR (microsatellite) markers. The lack of diversity detected among 105 cultivated and wild accessions of B. oleracea contrasted starkly with that found within its wild relatives. The vast majority of B. oleracea accessions shared a single haplotype, whereas as many as six haplotypes were detected in two wild species, B. villosa Biv. and B. cretica Lam.. The SSRs proved to be highly polymorphic across haplotypes, with calculated genetic diversity values (H) of 0.23-0.87. In total, 23 different haplotypes were detected in C genome species, with an additional five haplotypes detected in B. rapa L. (A genome n = 10) and another in B. nigra L. (B genome, n = 8). The low chloroplast diversity of B. oleracea is not suggestive of multiple domestication events. The predominant B. oleracea haplotype was also common in B. incana Ten. and present in low frequencies in B. villosa, B. macrocarpa Guss, B. rupestris Raf. and B. cretica. The chloroplast SSRs reveal a wealth of diversity within wild Brassica species that will facilitate further evolutionary and phylogeographic studies of this important crop genus.

Avaliação de espécies silvestres e cultivares de amendoim para resistência a Enneothrips flavens Moulton

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)