Methyl jasmonate and methyl salicylate, but not cis-jasmone, evoke defenses against infection of Arabidopsis thaliana by Orobanche aegyptiaca

The mechanism by which some plant species develop resistance to the root parasite, broomrape ( Orobanche aegyptiaca ), is still not clear. Resistance to other pathogens can be induced by methyl jasmonate and systemic acquired resistance can be induced by treatment with salicylic acid, while cis -jasmone can act as a signaling molecule in plant–insect interactions. The three compounds studied, methyl jasmonate, cis -jasmone, and methyl salicylate, were applied to Arabidopsis thaliana seedlings that were then transferred to Nunc cell culture plates and exposed to the germinating seeds of O. aegyptiaca . The number of infections of the roots of single seedlings of A. thaliana was then quantified. Exposure for 24 h to very low concentrations of methyl jasmonate or methyl salicylate, which were then removed, effectively induced resistance to infection of A. thaliana by O. aegyptiaca , reducing attachment and tubercle formation by 90%. cis -Jasmone was far less effective in inducing a similar resistance to infection. These results support the view that methyl jasmonate can induce almost full resistance to infection by broomrape. The fact that such resistance is not observed under normal conditions of infection supports the idea that the root parasite does not evoke the full defensive response in the host plant.     

Orobanche crenata resistance and avoidance in pea (Pisum spp.) operate at different developmental stages of the parasite

Orobanche crenata (broomrape) is an important constraint to pea (Pisum sativum) cultivation in the Mediterranean area, because little resistance is available in commercial crop varieties. Field experiments have demonstrated that some resistance is present in a number of P. sativum and P. fulvum accessions. The goal of this work was to characterize such resistance. The Pisum–O. crenata interaction and the resistance symptoms were studied under controlled conditions by using Petri dish and polyethylene bag assays. The content of phenolics and peroxidase activity in host tissue from infected and non-infected plants were also measured. Resistance and avoidance mechanisms, acting at different developmental stages of the parasite, have been identified, including low stimulation of O. crenata seed germination, unsuccessful penetration of host roots, delay in post-attachment tubercle development and necrosis of the attached tubercles. Infection caused an increase in the content of total soluble phenolics in some Pisum genotypes. Peroxidase activity was higher in resistant than in susceptible accessions. Results obtained with different Pisum genotypes showed that resistance is the result of several mechanisms acting at different stages of the infection process. Resistance is also related to increased levels of peroxidase activity in host roots.     

The role of peroxidase in the resistance of sunflower against Orobanche cumana in Russia

We present the results of a histochemical study comparing seedlings of races C and D of Orobanche cumana Wallr. (syn. O. cernua Loefl.) attacking sunflower (Helianthus annuus L.) in southern Russia. Three groups of O. cumana seedlings were distinguished according to the peroxidase content of the cells in the radicles: (1) those with neither extracellular nor intracellubr peroxidase and whose radicles have a smooth apex (these were classified as non-infective): (2) those with a high peroxidase content of the nuclei and the cytoplasm layer adjacent to the cell wall, as well as excretion of peroxidase from the apex of the radicles: (3) those with a similarly high peroxidase activity in the parasite cells, but without extracellular excretion. The apices of the radicles of the last two groups are swollen. It is suggested that these belong to O. cumana races C and D respectively. The extracellular peroxidase in O. cumana race C reacts with phenolic compounds, which are lignin precursors of the host, resulting in host resistance due to the formation of lignin layers in sunflowers possessing the Or3 gene for resistance. The absence of extracellular peroxidase in O. cumana race D prevents lignin formation and enables the parasite to attach to the host vascular system. Comparison of these data with the information on the earlier O. cumana races A and B, and older sunflower cultivars, points to a crucial role of peroxidase in the process of breeding new sunflower cultivars and the evolution of new O. cumana races.     

分枝列当寄生甜瓜时吸器形成过程中内源激素的动态变化

本研究采用石蜡切片和显微观察分析分枝列当寄生甜瓜时的吸器形成过程, 利用液相色谱-质谱联用 (LC-MS) 检测吸器形成过程中24种植物内源激素变化。结果表明:分枝列当种子在接种甜瓜根部第3 天开始萌发, 接种第7 天分枝列当吸器顶端稳固地附着在寄主根部, 吸器侵入甜瓜根皮层, 并在甜瓜根皮层细胞内扩展, 扩展过程中形成大量的爪状结构, 接种第11 天时分枝列当吸器细胞与寄主根维管束连接。共有17种植物激素参与了吸器形成过程, 其中生长素类吲哚-3-甲醛含量在列当吸器形成过程中降低了83.36%，细胞分裂素类反式玉米素核苷的含量在增长了5.02倍;油菜素内酯含量增长了481.95倍, 推测这3类植物激素在分枝列当吸器形成过程中可能起到重要的调控作用。     

Direct infection of potato tubers by the root parasite Orobanche aegyptiaca

Parasitic plants of the Orobanchaceae are known as obligate root parasites that develop haustoria that connect to roots of various host plants. This article describes, for the first time, a case where the root parasite successfully connected to potato tubers, i.e. to the swollen portion of an underground stem rather than to a root. The rhizosphere of Orobanche aegyptiaca and of its host Solanum tuberosum (potato) was carefully examined. In anatomical studies, the adventitious roots were directly connected to potato tubers. Numerous secondary haustoria, which developed along the adventitious roots in close vicinity to the potato tuber, penetrated the tuber epidermis and the perimedullary tuber parenchyma and developed xylem strands that are presumably connected to the minor xylem strands within the tuber cortex. These findings indicate that parasites of the Orobanchaceae that normally attack host roots may also parasitise underground stem tubers.     

A study of assimilation and translocation in Cuscuta hyalina Heyne ex Roth., Orobanche ramosa L. and Striga hermonthica Benth.

Quantitative determinations of chlorophyll showed that Cuscuta hyalina and Striga hermonthica possess chlorophyll. No trace of chlorophyll was found in Orobanche ramosa. Autoradiographic evidence showed that both radiocarbon and radiophosphorous moved from host to parasite in alt three species. Photosynthesis occurs in S. hermonthica when the parasite is isolated from its host. The adventitious roots of S. hermonthica and O. ramosa appear to be functional in absorbing a proportion of the required inorganic substances and water. S. hermonthica is a hemiparasite, O. ramosa is a holoparasite but C. hyalina is best described as a partial parasite.     

Primary Haustorial Development of Striga asiatica on Host and Nonhost Species

ABSTRACT Initial interactions of Striga asiatica with a susceptible host and non-host plants were examined by histological methods. Haustorial development was initiated when radicles of S. asiatica were placed in contact with host or nonhost roots. Reorganization of the S. asiatica root apical meristem was rapid and involved the formation of a distal group of cells that penetrated the host or nonhost root. Penetration of the epidermis of the host (sorghum) roots and advance into the cortex occurred within 24 to 48 h of inoculation. Penetration of the endodermis by the developing endophyte was delayed for 72 to 96 h after initial contact. However, upon penetration vascular continuity was established between parasite and host. In contrast, interactions with nonhosts provided evidence of active resistance mechanisms. Penetration of lettuce, marigold, and cowpea roots by S. asiatica was most frequently arrested in the cortex, and endophytic cells were necrotic 72 h after inoculation. Some species-specific differences were observed in the reactions of nonhosts to penetration, although in their general nature the interactions with S. asiatica were similar.     

Early stages of infection of maize (Zea mays) and Pennisetum setosum roots by the parasitic plant Striga hermonthica

Infection of young roots of maize (Zea mays L.) by the parasitic plant Striga hermonthica (Del.) Benth. was examined. Attachment to and penetration of roots occurred within 1–2 days after inoculation. Subsequent growth through the cortex to the host stele and proliferation of parasite xylem tissue was commonly completed by 3–4 days after inoculation. Histochemical staining showed that young maize roots do not contain major wall-thickening components. However, an increase in cell wall fluorescence and endodermal cell wall thickness was often seen at the site of infection and in the surrounding maize root tissue at 3 days after inoculation. This host response was variable and did not prevent rapid and successful penetration by the parasite. In contrast, uninfected roots of Pennisetum setosum (Sw.) L. Rich., a species resistant to S. hermonthica, had substantial thickening of the inner endodermal cell walls and exhibited further cell wall thickening at the stele upon infection. Examination of infections on both hosts demonstrated the presence of autofluorescent material at the host-parasite interface. This material was thicker and more extensive at the P. setosum-S. hermonthica interface than at the maize-S. hermonthica interface, and contained polyphenols and lignin. Examination of the host-parasite xylem connections in maize revealed substantial invasion of the host stele by both parenchyma and tracheary elements. In a few cases of P. setosum infection, parasite cells entered the stele; however, this did not lead to successful establishment of the parasite.     

Gene expression profiling of Medicago truncatula roots in response to the parasitic plant Orobanche crenata

Orobanche crenata is a root parasitic weed that is a major constraint for grain and forage legume cultivation in Mediterranean and West Asia. Only moderate to low levels of incomplete resistance of complex inheritance has been identified so far in legume crops, which has hampered genetic and genomic analysis. In the present study, we provide a gene expression profile of roots of the model legume Medicago truncatula in response to infection by O. crenata . M. truncatula accessions SA27774 (complete resistance acting at early penetration stages) and SA4087 (incomplete late acting resistance mediated by necrosis of parasite tubercle) were inoculated with O. crenata seeds in a semi-sterile dish system. Roots were harvested at 15 (first contacts of the parasitism structures with the host roots), 21 (initial stage of parasite tubercle formation on SA4087) and 35 (prior necrosis of well-developed parasite tubercle of on SA4087) days post-inoculation. Array hybridisations revealed several hundred genes up-regulated in response to O. crenata infection. Gene expression patterns suggest that resistance mechanisms activated in both genotypes are temporally and spatially different and resemble those associated with plant resistance to microbial pathogens. Regulated genes identified here represent a comprehensive resource that can be used as a support to breeding strategies for resistance.     

Defense enzymes induced in cucumber roots by treatment with plant growth-promoting rhizobacteria (PGPR) and Pythium aphanidermatum

Root and crown rot of cucumber caused by Pythium aphanidermatum can be suppressed by various rhizobacteria or PGPR (plant growth-promoting rhizobacteria). When cucumber roots were treated with Pseudomonas corrugata 13 or Pseudomonas aureofaciens 63–28, phenylalanine ammonia-lyase (PAL) activity was stimulated in root tissues in 2 days and this activated accumulation lasted for 16 days after bacterization. Peroxidase (PO) and polyphenol oxidase (PPO) activities were increased in roots 2–5 days after bacterization with P. corrugata strain 13. After bacterized cucumber roots were challenged with P. aphanidermatum, the enzyme activities of PAL, PO and PPO increased as the disease developed on the roots. These accumulations peaked 4–6 days after pathogen inoculation. A split root system demonstrated that the three enzymes could be systemically induced by the Pseudomonas strains 63–28 and 13, as well as P. aphanidermatum. Furthermore, isoperoxidase native PAGE (polyacrylamide gel electrophoresis) analysis indicated that the peroxidase isomer forms in cucumber roots induced by rhizobacteria were different from that in roots infected with P. aphanidermatum. These results suggest that the plant defense enzymes could be stimulated in cucumber roots which have been colonized by non-pathogenic rhizobacteria or in a compatible interaction between cucumber and P. aphanidermatum. The mechanisms of PO activation by the rhizobacteria may be different from those of pathogen infection.     

Some compatible Rhizobium leguminosarum strains in peas decrease infections when parasitised by Orobanche crenata

Orobanche crenata (crenate broomrape) produces serious damage to many legume crops and particularly becomes a limiting factor for pea production in the Mediterranean basin. Nodulation effects on pea–broomrape relationships were studied using the commercial pea cultivar Douce de Provence and different Rhizobium strains using pot and Petri dish experiments. First, the benefit of bacterial inoculation on plant growth and efficiency of N incorporation were demonstrated for two isolates, P.SOM and P.1236. These isolates did not influence parasite germination induced by the artificial stimulant, GR24. In contrast, pea root inoculation with P.SOM and P.1236 isolates led to a reduced root infection by O. crenata , resulting from a lower Orobanche germination rate close to pea roots and a limited capacity of the parasitic seedlings to develop tubercles. Broomrape necrosis was observed both before and after parasite attachment to inoculated pea roots. Concomitantly, reduction in infection was accompanied by enhanced peroxidase activity and constantly high phenylalanine ammonia lyase activity in pea roots. These data suggest the involvement of these enzymes in pea resistance to crenate broomrape induced by the compatible rhizobia. Management of Orobanche via crop selection based on these enzyme systems is a viable option.     

胡杨根际可培养细菌的分离及抑制瓜列当种子萌发研究

从胡杨生存的干旱强光照逆境中获得新的细菌资源,从中筛选新的可抑制瓜列当种子萌发的生防菌。采用3种分离培养基,以稀释涂布法对塔里木河流域3处胡杨根系样品进行可培养细菌分离,通过分析分离菌株16S rRNA基因序列确定其种属分类。采用皿内培养法测定其对瓜列当种子萌发的抑制活性,利用盆栽试验检测分离菌株M2-3对瓜列当寄生的防治效果。结果分离获得98株细菌,分属于28个属,马赛菌属Massilia、Pontibacter、玫瑰单胞菌属Roseomonas为优势菌属。其中,菌株M2-3与Pontibacter salisaro HMC5104T的相似度最高,为97.92%,其菌体发酵液10倍稀释液对瓜列当种子萌发抑制率为87.8%,盆栽试验表明该菌株对瓜列当防效可达36.9%,具备进一步开发可能性。     

Phytotoxic activity of root absorbed glyphosate in corn seedlings (Zea mays L.)

Growth chamber experiments were conducted in order to study the absorption, translocation and activity of glyphosate when applied to roots with aqueous solution avoiding any glyphosate–substrate interaction. Corn seedlings at the first leaf stage were set up in individual graduated cylinders containing different solutions of ¹⁴C-glyphosate (0–30 mg ae kg⁻¹). After 26 h of root exposure, plants were transferred to fresh nutrient solution and grown for the next 5 days. After harvest, plants were separated into seed, root, mesocotyle, coleoptile, cotyledon, first leaf and all new leaves (apex), and quantified ¹⁴C radioactivity contained in each part. Glyphosate uptake was only 11% of the theoretical mass flow into the plant. The amount of glyphosate translocated from roots was positively correlated with plant uptake ( P  < 0.01). Total plant fresh weight presented a logistic response to glyphosate amounts, including a growth stimulant effect (hormesis), when plants absorbed less than 0.6 µg. The treated plants presented a normal pattern of glyphosate allocation, with the apex the principal sink, accumulating more than 38% of mobilized glyphosate. When corn plants absorbed more than 0.6 µg they showed a decrease in growth. The relatively high glyphosate quantities allocated in the new leaves showed the relevance of the symplastic pathway in the translocation process for root absorbed glyphosate.     

Prohexadione-calcium induces sunflower (Helianthus annuus) resistance against the root parasitic weed Orobanche cumana

Prohexadione-calcium (PHDC) is a plant growth retardant that inhibits gibberellin biosynthesis and transiently alters the spectrum of flavonoids and their phenolic precursors. In this study, root chamber and pot experiments were conducted to evaluate the effects of PHDC on Orobanche cumana seed germination and induction of sunflower resistance to this root parasitic weed. Root chamber experiments indicated that PHDC did not interfere with the progress of Orobanche seed germination, but retarded O. cumana tubercle formation and development. In pot experiments, control of O. cumana by PHDC depended on its dose, method of application and sunflower cultivar. In sunflower cv. HA89, PHDC at 10 mg LS⁻¹ (litre soil) as two drenches or at 1 m m as two foliar sprays significantly reduced the total number and dry matter of O. cumana without influencing flower, leaf and root development. In sunflower cv. Albena, PHDC at 20 and 30 mg LS⁻¹, but not at 10 mg LS⁻¹, as two drenches significantly reduced O. cumana infection. Sunflower cv. HA89 was less susceptible to O. cumana and more inducible by PHDC than sunflower cv. Albena. The biochemical analysis of root extracts revealed a significantly higher accumulation of free phenolics in sunflower cv. HA89 after PHDC soil drench, but not in Albena. No significant lignification was observed after PHDC treatments. The results suggest that PHDC reduces O. cumana infection in sunflower by inducing host resistance, not by directly interfering with the parasite. Free phenolics play an important role in sunflower response to the parasite.     

An in-vitro growth system for studying the parasitism of cowpea (Vigna unguiculata) by Striga gesnerioides

An in-vitro growth system for investigating the behaviour of S. gesnerioides on cowpea has been developed. The roots of young cowpea plants were spread over glass fibre filter paper in a shallow plastic tray, and a known number of germinated Striga seeds were placed on the cowpea root surface. This allowed the infection process of the parasite to be quantified. Good access to the host-parasite association enabled the entire infection process to be easily monitored and tissue removed for cytological studies. Observations revealed for the first time that contact of S. gesnerioides radicles with host roots stimulated the development of radicular hairs and swelling of the tip of these radicles. Furthermore, penetration of cowpea roots by Striga was shown to be localized and to cause very little damage to the host roots. Swollen parasite tubercles developed on the surface of the roots only after the host stele had been penetrated by invading Striga cells. This growth system is suitable for in-vestigating the mechanisms of host resistance. and also for rapidly screening cowpea germplasm for new sources of resistance to the Parasite. Resistant cowpea cultivars offer the best potential solution for control of this Parasite.     

New insights concerning the ecology and the biology of Rhamphicarpa fistulosa (Scrophulariaceae)

Rhamphicarpa fistulosa (Hochst.) Benth. is a parasitic member of the Scrophulariaceae occurring sporadically on rice and other cereal crops in tropical Africa. Relatively little is known about this parasite, and some aspects concerning its life cycle and its impact on crops are reported in the present paper. The size, the shape and the seedcoat ornamentation of R. fistulosa are described and compared with those of Striga spp. The seeds do not require a host root exudate to stimulate germination but do require light. As in the case of Striga spp., they show a period of dormancy of at least 6 months. When the parasite roots come into close contact with a host root, they swell and develop haustoria, which form a direct xylem connection between host and parasite. In vitro and pot culture experiments establish that R. fistulosa is a facultative parasite, as it is able to complete its life cycle without a host plant. However, plants growing under these conditions are much smaller and produce only a few seeds. Field observations indicate that in some places, R. fistulosa can destroy cereals. The agronomic hazard represented by this member of the Scrophulariaceae is discussed.     

禾顶囊壳小麦变种对小麦种子根的侵染过程

 光镜和电镜观察表明,禾顶囊壳小麦变种(Gaeumannomyces graminis var.tritici,小麦全蚀病菌)对小麦种子根的侵染过程可分为侵入前、侵入表皮层、进入皮层和进入中柱等4个连续阶段。麦根接菌后在15℃下培养,48 h后侵入表皮层细胞,60 h后进入皮层,120 h后进入中柱。病原菌主要以侵染菌丝直接侵入表皮层,表皮细胞间隙和根毛基细胞是主要侵入部位,少数由附着枝侵入。菌丝穿透细胞壁有明显的酶解作用特征,菌丝先端前方胞壁上还产生电子密物质。皮层细胞是病原菌定殖和发展的主要场所,病原菌还能离解胞间层,形成胞外空间,特别有利于菌丝和菌丝束的扩展。在侵入位点的寄主细胞壁和质膜之间,形成多种形状的木质管,其数量与侵入菌丝的数目相对应,但木质管不能阻止菌丝进入细胞。菌丝进入中柱后,可阻塞导管和筛管。小麦细胞发生退行性病变,尤以细胞壁膨大崩坏和早期质壁分离最明显,细胞间隙还产生性质不明的黄色物质。     

Influence of environmental pH modulation on efficiency of apoplastic PR proteins during Fusarium culmorum– wheat seedling interaction

Variation of apoplastic pH by Fusarium culmorum and its influence on the production, activity and isoenzymes patterns of the pathogenesis‐related (PR) proteins β1,3‐glucanase, chitinase and peroxidase enzymes were detected in apoplastic fluids (AFs) from infected wheat seedlings. The time course in the 24–48 h interval post infection was characterized by an increase in activity and isoenzymatic differential induction of the selected PR proteins and by a concomitant rise of apoplastic pH. Chitinase attained maximum activity at pH 8·0 in the case of inoculated seedlings. Optimal β1,3‐glucanase activity in the pH range 6·0–8·0, was observed at pH 7·0. Peroxidase was strongly affected by pH, with enzyme activity having a maximum rate at pH 6·0 and thereafter rapidly declining at higher pH. Maximum peroxidase activity paralleled the appearance of the complete isoenzymatic pattern. In order to investigate the biological role of PR proteins in AFs, the in vitro antifungal activity was evaluated. In the interval 0–6 h, pH of macroconidia suspensions rose up to 7·2. AFs revealed inhibitory activity against germinating macroconidia of F. culmorum by decreasing the germination efficiency of macroconidia apical compartments, while this effect was compensated by an increased germination capacity of middle compartments. Present results suggest that during infection of wheat seedlings by F. culmorum the pH modulation favours host colonization by enhancing the activity of pectin lyase, and simultaneously inhibits the capacity of the host to oppose the pathogen by interfering with peroxidases which represent an important component of the defence arsenal.     

Initial Cellular Interactions Between Thielaviopsis basicola and Tobacco Root Hairs

ABSTRACT Cellular events that occur during the initial interactions between Thielaviopsis basicola and root hairs of tobacco (Nicotiana tabacum) were examined microscopically. Time-course documentation of the infection process indicated a dynamic interaction between T. basicola and the living host cell. Upon root hair contact and recognition, the vegetative apex of T. basicola rapidly differentiated to form infection structures, and the host cell responded cytologically. Penetration was achieved by threadlike hyphae that subsequently developed distal swellings, and intracellular hyphae of sickle-shaped morphology advanced from the distal swelling and colonized the cell. Streaming of the host cytoplasm became aggregated near the infection site prior to penetration and accumulated around the infecting hyphae as long as the host cell was viable. Substantial callose deposition, in the form of a bell-shaped collar around infection structures, resulted from the cytological activity at the infection site. Penetration of dead root hairs was common, but did not lead to the development of infection structures or to a sustained association with the host tissue; T. basicola exited dead root hairs and resumed vegetative growth. The establishment of the parasitic relationship by T. basicola was characteristic of hemibiotrophic fungi in that, initially, biotrophic infection led to tissue colonization, and host cell survival was limited under parasitism.     

Orobanche aegyptiaca control in tomato fields with sulfonylurea herbicides

Field experiments were conducted from 1994 to 1997 at two locations to study the effectiveness of chlorsulfuron and triasulfuron applied through different irrigation methods (chemigation) for control of Orobanche aegyptiaca Pers. in tomato ( Lycopersicon esculentum Mill). Three split applications of chlorsulfuron at 2.5 g a.i. ha⁻¹ and of triasulfuron at 7.5 g a.i. ha⁻¹, through conventional sprinkler irrigation systems, 10–14 days apart followed immediately by sprinkling with water, controlled O. aegyptiaca by about 90% and 80% and increased crop yield 25–47% and 30%, respectively, without any crop injury symptoms. Repeated applications of the same herbicides at half rates resulted in slightly higher O. aegyptiaca control and crop yield than only one herbicide application at double rate. Chemigation by the sprinkler systems (microsprinklers, 60 m³ ha⁻¹) slightly increased the herbicide efficiency as compared with the high volume spray (800 m³ ha⁻¹). O. aegyptiaca control from sulfonylureas applied by drip chemigation was poor, as this probably requires very accurate timing and the herbicide distribution in the soil was not uniform.