Germination stimulants of Phelipanche ramosa in the rhizosphere of Brassica napus are derived from the glucosinolate pathway

Phelipanche ramosa is a major parasitic weed of Brassica napus. The first step in a host-parasitic plant interaction is stimulation of parasite seed germination by compounds released from host roots. However, germination stimulants produced by B. napus have not been identified yet. In this study, we characterized the germination stimulants that accumulate in B. napus roots and are released into the rhizosphere. Eight glucosinolate-breakdown products were identified and quantified in B. napus roots by gas chromatography-mass spectrometry. Two (3-phenylpropanenitrile and 2-phenylethyl isothiocyanate [2-PEITC]) were identified in the B. napus rhizosphere. Among glucosinolate-breakdown products, P. ramosa germination was strongly and specifically triggered by isothiocyanates, indicating that 2-PEITC, in particular, plays a key role in the B. napus-P. ramosa interaction. Known strigolactones were not detected by ultraperformance liquid chromatography-tandem mass spectrometry, and seed of Phelipanche and Orobanche spp. that respond to strigolactones but not to isothiocyanates did not germinate in the rhizosphere of B. napus. Furthermore, both wild-type and strigolactone biosynthesis mutants of Arabidopsis thaliana Atccd7 and Atccd8 induced similar levels of P. ramosa seed germination, suggesting that compounds other than strigolactone function as germination stimulants for P. ramosa in other Brassicaceae spp. Our results open perspectives on the high adaptation potential of root-parasitic plants under host-driven selection pressures.     

An improved axenic system for studying pre-infection development of the parasitic plant Orobanche ramosa

BACKGROUND AND AIMS: Broomrapes (Orobanche spp.) are holoparasitic weeds that cause devastating losses in many economically important crops. The molecular mechanisms that control early stages of host infection in Orobanche are poorly understood, partly due to the lack of experimentally tractable in vitro systems that allow the efficient application of molecular tools. Here an improved axenic system for the analysis of pre-infection stages in O. ramosa in the absence of the host plant is described. METHODS: An optimized protocol for seed disinfection, based on formaldehyde, was developed. Orobanche ramosa seeds were conditioned in Petri dishes with filter paper, stimulated by addition of the synthetic strigol analogue GR24, and the percentage of germination as well as attachment-organ formation was determined. KEY RESULTS: Treatment of O. ramosa seeds with tobacco-root exudate or with GR24 resulted in highly reproducible germination rates around 70 %. A conditioning period of 8 d was both necessary and sufficient to allow optimal germination in response to GR24. Conditioned seeds that were dehydrated for several months remained fully responsive to GR24 without the need of a new conditioning period. Treatments as short as 5 min with GR24 were sufficient to fully and irreversibly induce the seed germination response. Approximately half of the germinated seeds initiated attachment-organ development. Similar rates of attachment organ induction were also detected in the rare cases of seeds that had germinated spontaneously on water. CONCLUSIONS: The results suggest that the conditioning period produces persistent changes in the seeds required for responsiveness to external stimulants. The rapid action of GR24 suggests that it may act via a receptor-mediated signalling mechanism. While germination in O. ramosa is induced by exogenous stimuli, attachment organ differentiation appears to be triggered by unknown endogenous signals. The new in vitro culture system will have useful applications for the molecular analysis of early stages of parasitic development in Orobanche.     

连续13年稻鸭共作兼秸秆还田的稻麦连作麦田杂草种子库物种多样性变化

稻鸭共作能有效控制稻田杂草的危害, 但是它对后茬小麦田杂草的影响及其控制作用尚没有详细的报道。我们于2000-2012年对江苏丹阳稻鸭共作兼秸秆还田的有机稻麦连作田土壤杂草种子库进行了连续13年的观察实验。结果显示, 稻鸭共作兼秸秆还田的措施使看麦娘(Alopecurus aequalis)、通泉草(Mazus japonicus)、碎米荠(Cardamine hirsuta)等18种麦田主要杂草的种子库均有较大幅度的降低, 总体的降低幅度高达97%。除了Pielou指数处于小幅波动状态外, 麦田杂草群落多样性指数整体呈下降趋势。丰富度下降表明杂草种子库向种类少、多样性低的方向演变。从Bray-Curtis指数和Jaccard相似性指数也可以得到同样的结论。可见, 连续稻鸭共作兼秸秆还田能够降低下茬的麦田土壤里杂草种子密度及多样性, 控制杂草危害。     

Stimulation of Orobanche ramosa seed germination by fusicoccin derivatives: a structure-activity relationship study

A structure-activity relationship study was conducted assaying 25 natural analogues and derivatives of fusicoccin (FC), and cotylenol, the aglycone of cotylenins, for their ability to stimulate the seed germination of the parasitic species Orobanche ramosa. Some of the compounds tested proved to be highly active, being 8,9-isopropylidene of the corresponding FC aglycone and the dideacetyl derivative the most active FC derivatives. In both groups of glucosides and aglycones (including cotylenol), the most important structural feature to impart activity appears to be the presence of the primary hydroxy group at C-19. Furthermore, the functionalities and the conformation of the carbotricyclic ring proved to play a significant role. The dideacetyl derivative of FC, being easily and rapidly obtainable in high yield starting by FC, could be of interest for its practical application as a stimulant of Orobanche ramosa seed germination, inducing the "suicidal germination", an interesting approach for parasitic plant management.     

IAA production during germination of Orobanche spp. seeds

Broomrapes (Orobanche spp.) are parasitic plants, whose growth and development fully depend on the nutritional connection established between the parasite and the roots of the respective host plant. Phytohormones are known to play a role in establishing the specific Orobanche-host plant interaction. The first step in the interaction is seed germination triggered by a germination stimulant secreted by the host-plant roots. We quantified indole-3-acetic acid (IAA) and abscisic acid (ABA) during the seed germination of tobacco broomrape (Orobanche ramosa) and sunflower broomrape (O. cumana). IAA was mainly released from Orobanche seeds in host-parasite interactions as compared to non-host-parasite interactions. Moreover, germinating seeds of O. ramosa released IAA as early as 24 h after the seeds were exposed to the germination stimulant, even before development of the germ tube. ABA levels remained unchanged during the germination of the parasites' seeds. The results presented here show that IAA production is probably part of a mechanism triggering germination upon the induction by the host factor, thus resulting in seed germination.     

稻田水面漂浮的杂草种子种类及动态

为了揭示杂草种子的漂浮动态,并为探寻生态控草方法提供理论依据,我们于2005年6-11月,在南京沧波门稻作区调查了水稻种植前及生长后期田问杂草群落以及稻田灌水期间水面漂浮的杂草种子的种类及漂浮时间.结果表明,在水稻种植前及生长后期田间出现有15科24种杂草;可漂浮于水面的杂草种子有17科26种,与田间发生的杂草种类相同的共有18种,占所有32种杂草的56.25%.这些杂草主要隶属于禾本科、蓼科、毛茛科、报春花科、莎草科、菊科和雨久花科.田埂附近漂浮的杂草种类及种子数量均高于田中间;各类杂草种子的漂浮能力不一,其中菵草(Beckmannia syzigachne)、泽珍珠菜(Lysimachia candida)、茴茴蒜(Ranunculus chinensis)、齿果酸模(Rumex dentatus)、蛇床(Cnidium monnieri)、鳢肠(Eclipta prostrata)和看麦娘(Alopecurus aequalis)等在田间水面漂浮能力较强.漂浮期间杂草种子除多集中于田埂附近,也附着于稻稞周围,并有互相结联的趋势.从整体上看,杂草种子漂浮2天后骤降,8天后基本全部沉降.显然,稻田生境的杂草种子大多可以漂浮于水面传播扩散.因此在灌水早期采取过滤或清除水上漂浮杂草种子可以减少杂草种子的输入和加速杂草种子从田间输出,减少杂草发生基数.     

Recovery of native grass biodiversity by sowing on former croplands: Is weed suppression a feasible goal for grassland restoration?

Grassland restoration on former croplands offers good opportunity to mitigate the loss of grassland biodiversity. Weed suppression can be another benefit, which becomes increasingly important because of the high recent rate of abandonment of arable lands in Central and Eastern Europe. Our aim was to evaluate the usefulness of sowing two low-diversity seed mixtures followed by annual mowing, a frequently used restoration technique, in weed suppression. We found that rapidly forming cover of sown grasses effectively suppressed short-lived weeds and their germination except in the first year. The detected dense seed bank of short-lived weeds points out the possibility and threat of later weed infestation. In the short run perennial weeds cannot be suppressed easily by sowing and annual mowing. We found that the effectiveness of seed sowing followed by mowing in weed suppression can be different on sites with different history or seed mixture. Rapidly establishing perennial weeds, such as Agropyron species were only detected in former alfalfa fields; Cirsium arvense was found in former cereal and sunflower fields but not in former alfalfa fields. We found that the rate of weed suppression and success was influenced by the seed mixtures used. In several alkali restorations the high proportion of perennial weeds was detected in year 3. In loess restorations, much lower scores were typical. This was likely caused by the different seed mixture used. The loess seed mixture contained seeds of a clonally spreading tall-grass, Bromus inermis, which could compete more effectively with clonally spreading weeds, than could short grass species with or without tussock forming. Our findings indicate that post-restoration management require carefully designed actions that are fine-tuned addressing specific threats at the site level.     

The conservation of arable weeds at crop edges of barley fields in northeast Spain

The effects of weed control practices and fertilisation on weed flora and crop yield were evaluated in crop edges of barley fields in northeastern Spain. The study was carried out in four organic and four conventional barley fields. In each field, four permanent plots were delimited at the crop edge, and fertilisation and weed control treatments in a factorial design were applied over 3 years. Weed composition and the aboveground biomass of weeds and barley were recorded before the crop harvest in the first and the third year. We found relatively low values of species richness per field, as well as low values of weed biomass, especially in the organic crop edges (3.9% of total biomass). Weeds were significantly reduced by herbicide applications on conventional fields and were not affected by weed harrowing on organic fields or fertilisation. These results demonstrate that specific measures are needed to enhance biodiversity at crop edges both in organic and conventional fields. Our results also suggest that under Mediterranean conditions and among impoverished weed communities, limiting the use of herbicides is crucial to enhancing arable diversity and that, contrary to findings found in previous studies in temperate climates, fertilisation and weed harrowing have little effect on weeds.     

Variability of interactions between barrel medic (Medicago truncatula) genotypes and Orobanche species

A number of broomrapes ( Orobanche spp.) are major threats for grain legumes in the Mediterranean area. Previous studies have shown very high levels of resistance to Orobanche crenata in Medicago truncatula with little variation among accessions hampering, therefore, its use for genetic analysis. In order to identify alternative systems for ulterior genetic and genomic analysis, we studied early stages of the interaction between M. truncatula accessions and a range of Orobanche species. We found significant differences in the induction of germination on Orobanche aegyptiaca , Orobanche foetida var. broteri , Orobanche minor , Orobanche nana and Orobanche ramosa seeds. Accessions also varied in the number of O. aegyptiaca , O. crenata , O. foetida var. broteri , O. nana and O. ramosa attachments supported. Applications of the synthetic germination stimulant GR24 increased the germination level of Orobanche cumana and O. minor . No attachments were observed for any of these species because of physical barriers typical of host resistance. On the contrary, increase of O. nana germination by GR24 led to an increase in number of attachments albeit the normal development of the nodules was stopped in a later stage. The genetic variation observed for induction of germination and subsequent attachment will be useful for isolating and characterising genes involved in early stages of Orobanche –host plant interaction and for the study of the biosynthetic pathways of production of germination stimulants.     

Alternate paths of evolution for the photosynthetic gene rbcL in four nonphotosynthetic species of Orobanche

We have determined the nucleotide sequence for the Rubisco large subunit from four holoparasitic species of Orobanche. Intact open reading frames are present in two species (O. corymbosa and O. fasciculata), whereas the remaining species (O. cernua and O. ramosa) have rbcL pseudogenes. Sequences for rbcL 5'-UTRs from species of Orobanche have few changes in the promoter and ribosome binding sites compared to photosynthetic higher plants. Comparison of rbcL 3'-UTR sequences for Nicotiana, Ipomoea, Cuscuta, and Orobanche reveal that nucleotide sequences from parasitic plants have regions capable of forming stem-loop structures, but 56–69 nt are deleted upstream of the stem-loop in the parasitic plants compared to their photosynthetic relatives. Although rbcL pseudogenes of O. cernua and O ramosa have many large and small deletions, few indels are shared in common, implying that their common ancestor probably had an intact rbcL reading frame. Intact rbcL reading frames in O. corymbosa and O. fasciculata retain a bias of synonymous over nonsynonymous substitutions and deduced protein sequences are consistent with potentially functional Rubisco large subunit proteins. A conservative model of random substitution processes in pseudogene sequences estimates that the probability is low (P<0.028) that these sequences would retain an open reading frame by chance. Species of Orobanche have either had recent photosynthetic ancestors, implying multiple independent losses of photosynthesis in this genus, or the rbcL gene may serve an unknown function in some nonphotosynthetic plants.     

应用主成分分析法和最小生成树法研究浙中秋旱作物田杂草种间生态关系

研究意义浙江金华地处浙江中部,旱地面积有５．０９×１０４ｈａ,占耕地总面积的１８％以上,棉花、大豆、甘薯等秋旱作物是该地区重要的经济作物,几年来,由于农村劳动力转移,中耕除草减少,草害问题日趋严重。化学除草是解决旱田草害的有效方法。但是,除草剂的长期...     

Exotic wildland weeds serve as reservoirs for a newly introduced cole crop pest,Bagrada hilaris (Hemiptera: Pentatomidae)

The role of wildland weeds that serve as alternate hosts for insect pests has not been directly examined for the potential to sustain pest populations or facilitate pest outbreaks. The development of weed biological control programmes is also complicated by weed species that are closely related to economically important or native plants, especially rare or special status species. In recent surveys in southern California, USA, we found a newly introduced insect pest of cole crops, Bagrada hilaris Burmeister (Bagrada bug), feeding on Brassicaceae weeds in riparian areas adjacent to agricultural fields where cole crops are routinely grown. Insect populations grew to levels well over action thresholds and caused severe damage to populations of the invasive weed, Lepidium latifolium (perennial pepperweed). The numerical response of B. hilaris on L. latifolium and other Brassicaceae weeds in natural areas may pose a significant challenge to effectively managing pest populations in crops. However, the accidental introduction of this insect provides the opportunity to examine plant–insect interactions with important implications for development of biologically based control methods for weeds.     

安徽沿江圩丘农区夏收作物田杂草群落分布规律的研究

作者创用了七级目测调查取样,进行数量分析的方法,对夏收作物田杂草群落进行分类研究,揭示了安徽沿江圩丘农区夏收作物田杂草群落的分布规律主要受土壤类型和轮作制度制约。看麦娘杂草群落分布于水稻土田,猪殃殃,野燕麦杂草群落分布于丘岗区旱地,卷耳、婆婆纳杂草群落分布于灰潮土旱地。首次提出综合草害指数,用以评定杂草群落类型中每种杂草的危害性,从而,提出了不同土壤类型和轮作制度的田块中恶性杂草的定量化指标与防除策略。     

江苏南部麦田杂草群落发生分布规律的数量分析

利用七级目测法对江苏省南部麦田81个样点796块田地共计82,7hm^2草害进行调查,采集数据,经转化后,对其进行主成分(PCA)和系统聚类分析,并赋予生态学意义的解释,研究表明,决定江苏省南部麦田杂草的发生、危害和分布的主要因素是种植制度导致的田间土壤饱和水分持续时间和土壤酸碱度,生境相似,杂草群落具有趋同性,并就杂草发生的特点提出杂草综合治理的建议。     

长期不同施肥方式对麦田杂草群落的影响

以太湖地区农业科学研究所31a的长期肥料定位试验田为材料,分别于2011—2012年小麦苗期、拔节期和收获期进行了杂草群落调查,研究杂草类型与密度的分布、杂草多样性指数的变化,并对杂草种群分布与土壤养分因子进行冗余分析。结果表明:小飞蓬、看麦娘、大巢菜、稻槎菜、通泉草是本地区小麦生长期的主要杂草类型;随着小麦的生长以及氮肥、有机肥的施入,杂草密度呈下降趋势;施入有机肥降低了麦季杂草的群落多样性指数,在小麦生长的不同时期均衡施肥的CNPK处理以及不施肥的C0处理的群落各项多样性指数能维持在一个较高的水平。RDA分析显示土壤氮含量以及有机质含量与第一排序轴相关性大,是对杂草分布影响最大的两个土壤养分因子。太湖地区稻麦两熟制条件下,长期有机无机肥料单一或配合投入可显著影响麦田杂草的群落组成,其中氮肥和有机肥的施入能显著降低杂草密度;土壤养分的差异影响田面杂草密度和优势种群,均衡施肥能降低优势杂草种群的优势地位,抑制其发生危害程度,提高农田生态系统的生产力及稳定性。     

种植制度对江苏省棉田杂草群落影响的研究

 经连续5年对江苏省主棉区水旱轮作及旱连作两种种植制度下的棉田杂草种群密度及草害优势度级数进行定田调查,通过时间和空间上的比较统计分析,研究了棉田杂草群落演替趋势及草害发生规律。研究结果表明,种植制度与江苏省棉田杂草群落的组成和草害发生程度紧密相关。在水旱轮作棉田中,随着棉花(Gossypium hirsutum)种植年数的增加,喜湿性杂草稗草(Echinochloa crusgalli)等优势度值逐渐减少,喜旱性杂草马唐(Digitaria sanguinalis)等优势度值逐渐提高,它们之间存在显著的线性关系。旱连作棉田以喜旱性杂草为主,发生量较大,草害较重,年际间杂草群落变化较小。另外,水旱轮作和旱连作棉田之间的杂草群落结构和种群数量差异显著。而地理区域差异的影响明显小于轮作制度差异导致的影响。     

Searching for the origins of arable weeds in the Near East

This short note adds to earlier attempts at identifying arable weeds on late Pleistocene/early Holocene sites in the Near East. Nineteen potential arable weed taxa that have no known use were selected. The occurrence of these taxa at sites with morphologically wild cereals was compared to sites with morphologically domestic cereals. The presumed arable weed taxa were as common on three PPNA (Pre Pottery Neolithic A) sites without domestication as they were on Middle PPNB (Pre Pottery Neolithic B) sites with domestication, which lends support to arguments for pre-domestic cultivation at the former sites. Arable weed taxa were less common at Natufian sites but their presence raises the question of whether they originated in cultivated fields or were the ancestors of weeds gathered accidentally alongside wild cereals in their natural habitat.