Biological and Transcriptomic Characterization of Pre-Haustorial Resistance to Sunflower Broomrape (Orobanche cumana W.) in Sunflowers (Helianthus annuus)

Infestations with sunflower broomrape (Orobanche cumana Wallr.), an obligatory root parasite, constitute a major limitation to sunflower production in many regions around the world. Breeding for resistance is the most effective approach to reduce sunflower broomrape infestation, yet resistance mechanisms are often broken by new races of the pathogen. Elucidating the mechanisms controlling resistance to broomrape at the molecular level is, thus, a desirable way to obtain long-lasting resistance. In this study, we investigated broomrape resistance in a confectionery sunflower cultivar with a robust and long-lasting resistance to sunflower broomrape. Visual screening and histological examination of sunflower roots revealed that penetration of the broomrape haustorium into the sunflower roots was blocked at the cortex, indicating a pre-haustorial mechanism of resistance. A comparative RNA sequencing between broomrape-resistant and -susceptible accessions allowed the identification of genes that were significantly differentially expressed upon broomrape infestation. Among these genes were β-1,3-endoglucanase, β-glucanase, and ethylene-responsive transcription factor 4 (ERF4). These genes were previously reported to be pathogenesis-related in other plant species. This transcriptomic investigation, together with the histological examinations, led us to conclude that the resistance mechanism involves the identification of the broomrape and the consequent formation of a physical barrier that prevents the establishment of the broomrape into the sunflower roots.

Costunolide Influences Germ Tube Orientation in Sunflower Broomrape – A First Step Toward Understanding Chemotropism

Orobanche cumana WALLR. is a host-specific root parasite of cultivated sunflowers with increasing economic importance in Europe, North Africa, and parts of Asia. While sesquiterpene lactones (STLs) released from sunflower roots were identified as natural germination stimulants of O. cumana seeds in the soil, the chemical nature of the signals guiding the emerging germ tube toward the host root has remained unknown hitherto. Thus, we designed a bioassay that allowed the observation of broomrape germination and subsequent germ tube development in the presence of substances with putative chemotropic activity. Root exudates and sunflower oil extracts, both containing STLs in micromolar concentrations, caused the positive chemotropic orientation of germ tubes. A similar positive chemotropic effect was achieved with costunolide, one of the four STLs of sunflower present in the exudate and oil extracts. In contrast, GR24, a synthetic strigolactone (SL) with germination-inducing activity on O. cumana seeds, showed no effect on the germ tube orientation. The effect of costunolide was concentration-dependent and within the range of its natural micromolar occurrence in roots. We assume that an STL gradient is responsible for the stronger inhibition of elongation growth on the host-facing flank of the germ tube compared with the far side flank. This would confer a double role of STLs from sunflower root exudates in the sunflower–broomrape interaction, namely, as germination stimulants and as chemotropic signals.

On the rediscovery of a rare root parasite Gleadovia ruborum Gamble & Prain (Orobanchaceae) from Uttarakhand, western Himalaya, India

Gleadovia ruborum Gamble & Prain (Orobanchaceae), a rare root parasite is recorded after a gap of 57 years from a new locality in Uttarakhand, western Himalaya. A total of four individuals were located in a shady moist forest of Abies pindrow-Quercus floribunda at Surkanda hill near Mussoorie. All the individual plants were growing on the roots of Rubus pedunculosus D. Don. The current communication provides an updated description, habitat characteristics and host species of Gleadovia ruborum.

Nitrates increase abscisic acid levels to regulate haustoria formation in the parasitic plant Phtheirospermum japonicum

Parasitic plants are globally prevalent pathogens that withdraw nutrients from their host plants using an organ known as the haustorium. Some, the obligate parasites are entirely dependent on their hosts for survival, whereas others, the facultative parasites, are independent of their hosts and infect depending on environmental conditions and the presence of the host. How parasitic plants regulate their haustoria in response to their environment is largely unknown. Using the facultative root parasite Phtheirospermum japonicum, we found that external nutrient levels modified haustorial numbers. This effect was independent of phosphate and potassium but nitrates were sufficient and necessary to block haustoria formation. Elevated nitrate levels prevented the activation of hundreds of genes associated with haustoria formation, downregulated genes associated with xylem development and increased levels of abscisic acid (ABA). Enhancing ABA levels independently of nitrates blocked haustoria formation whereas reducing ABA biosynthesis allowed haustoria to form in the presence of nitrates suggesting that nitrates mediated haustorial regulation in part via ABA production. Nitrates also inhibited haustoria formation and reduced infectivity of the obligate root parasite Striga hermonthica, suggesting a more widely conserved mechanism by which parasitic plants adapt their extent of parasitism according to nitrogen availability in the external environment.

Orobanche cumana (sunflower broomrape).

O. cumana is an obligatory, non-photosynthetic root parasite. It is believed to have evolved relatively recently from forms of O. cernua parasitizing wild Asteraceae, in particular species of Artemisia, and transferring to cultivated Helianthus annuus (sunflower). O. cumana is thought to be native to Eastern Europe (Russia) and has subsequently spread to most other sunflower growing regions of central and western Europe and Asia. The absence of O. cumana in sunflower growing regions of South America (for example Argentina) is believed to be associated with warmer winter temperatures not suitable for this species, rather than the seeds not being present. O. cumana can cause immense damage to cultivated sunflowers resulting in a significant decrease in yield. Despite resistant sunflower varieties being developed more virulent races of O. cumana have repeatedly evolved, or been selected, to overcome resistance. Thus, in spite of constant breeding efforts, losses continue in established sunflower growing areas and there is potential for it to invade new areas, wherever sunflower is grown.

Orobanche cernua (nodding broomrape).

O. cernua is an obligatory, non-photosynthetic root parasite which is native over a wide range across northeast Africa, southern Europe and western and southern Asia. In many of these areas it is a serious pest of Solanaceaeous crops such as Solanum lycopersicum (tomato) Nicotiana tabacum (tobacco) and S. melongena (aubergine) and occasionally S. tuberosum (potato). Species of Orobanche depend totally on their hosts for all nutrition and become an active sink for the host plant. This therefore results in a decrease in crop yield and as a result can have a major impact on the economy and livelihoods. Once established, the seed bank may last 10-20 years and there are no simple, economic control measures. Seeds of O. cernua are very small and inconspicuous and can be accidentally introduced into new areas as a contaminant of soil, seeds and machinery. There is potential for this species to invade many other areas of the world.

Biological and Transcriptomic Characterization of Pre-haustorial Resistance to Sunflower Broomrape (Orobanche cumana W.)

Infestations with sunflower broomrape (Orobanche cumana Wallr.), an obligatory root parasite, constitute a major limitation to sunflower production in many regions around the world. Breeding for resistance is the most effective approach to reduce sunflower broomrape infestation, yet resistance mechanisms are often overcome by new races of the pathogen. Elucidating the mechanisms controlling the resistance to broomrape at the molecular level is thus the most desirable pathway to obtaining long-lasting resistance and reducing yield loss in sunflower. In this study, we investigated broomrape resistance in a confectionery sunflower hybrid with a robust and long-lasting resistance to sunflower broomrape. Visual screening and histological examination of sunflower roots revealed that penetration of the intrusive broomrape cells into the host root endodermis is blocked at the host cortex, indicating a pre-haustorial mechanism of resistance. A comparative RNA-Seq experiment conducted between roots obtained from the resistant cultivar, a bulk of five broomrape resistant lines and a bulk of five broomrape susceptible lines allowed the identification of genes that were significantly differentially expressed upon broomrape infestation. Among these differentially expressed genes, β-1,3-endoglucanase, β-glucanase and ethylene-responsive transcription factor4 (ERF4) genes were identified. These genes were previously reported to be pathogenesis-related genes in other plant species. This genetics investigation together with the histological examinations led us to conclude that the resistance mechanism involves the identification of the broomrape and the consequent formation of a physical barrier that prevents the penetration of the broomrape into the sunflower roots.

First report of the root parasite Cansjera rheedei (Santalales: Opiliaceae) in Taiwan

The family Opiliaceae in Santalales comprises approximately 38 species within 12 genera distributed worldwide. In Taiwan, only one species of the tribe Champereieae, Champereia manillana, has been recorded. Here we report the first record of a second member of Opiliaceae, Cansjera in tribe Opilieae, for Taiwan. The newly-found species, Cansjera rheedei J.F. Gmelin (Opiliaceae), is a liana distributed from India and Nepal to southern China and western Malaysia. This is the first record of both the genus Cansjera and the tribe Opilieae of Opiliaceae in Taiwan. In this report, we provide a taxonomic description for the species and colour photographs to facilitate identification in the field.

Taxonomic traits in the microstructure of flowers of parasitic Orobanche picridis with particular emphasis on secretory structures

Orobanche picridis is an obligate root parasite devoid of chlorophyll in aboveground organs, which infects various Picris species. Given the high level of phenotypic variability of the species, the considerable limitation of the number of taxonomically relevant traits (mainly in terms of generative elements), and the low morphological variation between species, Orobanche is regarded as one of the taxonomically most problematic genera. This study aimed to analyse the taxonomic traits of O. picridis flowers with the use of stereoscopic and bright-field microscopy as well as fluorescence, scanning, and transmission electron microscopy. The micromorphology of sepals, petals, stamens, and pistils was described. For the first time, the anatomy of parasitic Orobanche nectaries and the ultrastructure of nectaries and glandular trichomes were presented. Special attention was paid to the distribution and types of glandular and non-glandular trichomes as well as the types of metabolites contained in these structures. It was demonstrated that the nectary gland was located at the base of the gynoecium and nectar was secreted through modified nectarostomata. The secretory parenchyma cells contained nuclei, large amyloplasts with starch granules, mitochondria, and high content of endoplasmic reticulum profiles. Nectar was transported via symplastic and apoplastic routes. The results of histochemical assays and fluorescence tests revealed the presence of four groups of metabolites, i.e. polyphenols (tannins, flavonoids), lipids (acidic and neutral lipids, essential oil, sesquiterpenes, steroids), polysaccharides (acidic and neutral polysaccharides), and alkaloids, in the trichomes located on perianth elements and stamens.