Micromorphological studies on seeds of orobanche species from the iberian peninsula and the balearic islands, and their systematic significance

BACKGROUND AND AIMS: Previous research has made clear the intrinsic taxonomic difficulties in identifying species in the genus Orobanche. The aim of this study, therefore, was to investigate the systematic utility of seed characteristics. METHODS: Light and scanning electron microscopy was used to examine the seeds of 33 taxa of Orobanche from the Iberian Peninsula and the Balearic Islands. KEY RESULTS: Characters such as size, shape and ornamentation of the seeds were not found to be very useful in differentiation of taxa; however, other characters of the epidermal seed coat cells proved to be very helpful in this respect. Ornamentation of the periclinal walls could be used to discriminate four morphological types. Other features related to the anticlinal walls of the cells, such as thickness, presence/absence of a narrow trough, or relative depth, all contributed to the characterization of a large number of species. CONCLUSIONS: The usefulness of micromorphological studies on seeds of Orobanche in relation to differentiating taxa is demonstrated, and a key is provided to distinguish species or groups of species.     

A peptide from insects protects transgenic tobacco from a parasitic weed

Parasitic plants present some of the most intractable weed problems for agriculture in much of the world. Species of root parasites such as Orobanche can cause enormous yield losses, yet few control measures are effective and affordable. An ideal solution to this problem is the development of parasite-resistant crops, but this goal has been elusive for most susceptible crops. Here we report a mechanism for resistance to the parasitic angiosperm Orobanche based on expression of sarcotoxin IA in transgenic tobacco. Sarcotoxin IA is a 40-residue peptide with antibiotic activity, originally isolated from the fly, Sarcophaga peregrina. The sarcotoxin IA gene was fused to an Orobanche-inducible promoter, HMG2, which is induced locally in the host root at the point of contact with the parasite, and used to transform tobacco. The resulting transgenic plants accumulated more biomass than non-transformed plants in the presence of parasites. Furthermore, plants expressing sarcotoxin IA showed enhanced resistance to O. aegyptiaca as evidenced by abnormal parasite development and higher parasite mortality after attachment as compared to non-transformed plants. The transgenic plants were similar in appearance to non-transformed plants suggesting that sarcotoxin IA is not detrimental to the host.     

Generation of transgenic asulam-resistant potatoes to facilitate eradication of parasitic broomrapes (Orobanche spp.), with the sul gene as the selectable marker

 Root-attaching parasitic flowering broomrapes (Orobanche spp.) are major constraints to vegetable, legume and sunflower production around the Mediterranean and elsewhere, with banned methyl-bromide fumigation or land abandonment of these affected crops as the major “solutions” to the problem. We report the specific generation of transgenic asulam-resistant potatoes as a way to eradicate this pest. The target-site resistance in the crop allows the herbicide to be translocated from treated leaves to the parasite via crop roots. This inhibitor of dihydropteroate synthase lethally then prevents folic acid biosynthesis in the parasite. Additionally, we demonstrate that asulam can be used directly in selecting resistant transformants, without the need for another selectable marker. Received: 18 June 1997 / Accepted: 17 July 1997     

Fabacyl acetate, a germination stimulant for root parasitic plants from Pisum sativum

A germination stimulant, fabacyl acetate, was purified from root exudates of pea (Pisum sativum L.) and its structure was determined as ent-2′-epi-4a,8a-epoxyorobanchyl acetate [(3aR,4R,4aR,8bS,E)-4a,8a-epoxy-8,8-dimethyl-3-(((R)-4-methyl-5-oxo-2,5-dihydrofuran-2-yloxy)methylene)-2-oxo-3,3a,4,5,6,7,8,8b-decahydro-2H-indeno[1,2-b]furan-4-yl acetate], by 1D and 2D NMR spectroscopic, ESI- and EI-MS spectrometric, X-ray crystallographic analyses, and by comparing the ¹H NMR spectroscopic data and relative retention times (RR_t) in LC-MS and GC-MS with those of synthetic standards prepared from (+)-orobanchol and (+)-2′-epiorobanchol. The ¹H NMR spectroscopic data and RR_t of fabacyl acetate were identical with those of an isomer prepared from (+)-2′-epiorobanchol except for the opposite sign in CD spectra. This is the first natural ent-strigolactone containing an epoxide group. Fabacyl acetate was previously detected in root exudates of other Fabaceae plants including faba bean (Vicia faba L.) and alfalfa (Medicago sativa L.).     

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.     

列当寄生对不同品种向日葵幼苗生长及抗氧化酶活性的影响

采用连作重茬含向日葵列当的土壤,进行不同列当抗性向日葵品种苗期盆栽试验,调查播种后第30、50和70天向日葵幼苗根系列当的寄生率、幼苗地上部和地下部干物质量及株高,并测定其叶片丙二醛(MDA)、苯丙氨酸解氨酶(PAL)、超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活性。结果表明:不同向日葵品种苗期对列当抗性明显不同,其中油葵品种‘T012244’和‘MGS’对向日葵列当寄生免疫,‘S31’和‘白杂9号’为高抗,而‘星火大白边’和‘白杂6号’为敏感。列当寄生后明显抑制敏感品种向日葵幼苗生长,而对免疫品种和高抗品种的影响较小。列当寄生后的敏感性品种‘星火大白边’幼苗叶片MDA含量迅速上升,PAL活性也大幅度增加,列当对幼苗生长伤害加重;而苗期免疫品种"T012244"和高抗品种"S31"MDA含量下降,PAL活性则变化不大。随接种时间延长,不同抗性向日葵品种苗期SOD、POD和CAT活性均呈现先升高后降低的变化趋势,且免疫品种的保护酶活性变化幅度大于敏感品种,说明列当寄生对敏感向日葵品种造成了逆境胁迫,诱导其幼苗叶片保护酶系统产生防御响应。     

Responsiveness of Orobanche ramosa L. Seeds to GR 24 as Related to Temperature, Oxygen Availability and Water Potential During Preconditioning and Subsequent Germination

Broomrape (Orobanche ramosa L.) is a root holoparasite responsible for important yield losses in numerous crops, particularly in the Mediterranean area. In this paper, the effects of temperature, oxygen concentration and water potential of the medium on broomrape seed germination were investigated. Seeds became able to germinate in the presence of a strigol analogue (GR 24) only after a preincubation period for at least 3 days at 20 °C. Their responsiveness to GR 24 increased with increasing duration of their preconditioning at 20 °C, and was optimal after 2–3 weeks. The preconditioning treatment was effective at temperatures ranging from 10 to 30 °C. At the optimal temperature (20 °C), it required at least 1% oxygen in the atmosphere and remained effective at a water potential of the medium of –2 MPa. A too prolonged preincubation of seeds at sub- or supraoptimal temperatures (5 and 30 °C) resulted in induction of a secondary dormancy. Seeds preconditioned for 14 days at 20 °C germinated in the presence of 1 mg L^–1 GR 24 at temperatures ranging from 10 to 25 °C, and the thermal optimum was the same (20 °C) than that of preconditioning. At 20 °C, seeds were able to germinate in the presence of GR 24 under atmospheres containing at least 3% oxygen and at a water potential of the medium as low as –3 MPa. The differences observed in the effects of environmental factors on preconditioning efficiency and germination of preconditioned seeds suggest that both processes involve different mechanisms. The results obtained might also help to better understand the regulation of O. ramosa spread in temperate areas.     

中草药浸提液对小列当种子发芽的影响

选用240种常用中草药,研究其蒸馏水和甲醇浸提液原液及其10倍、100倍和1 000倍稀释液对小列当种子发芽的影响.结果表明:小列当种子在蒸馏水浸提液刺激下发芽率超过70%的中草药有灯心草、降香和莲须,在甲醇浸提液刺激下发芽率超过70%的中草药有百部、桑椹、谷芽和萆薢;蒸馏水浸提液对小列当种子发芽抑制率超过50%的中草药有菊花、棕榈、苦杏仁和黄柏,甲醇浸提液对小列当种子发芽抑制率超过70%的有续断、地黄、肉豆蔻、地肤子、黄连、木贼和胡黄连.大部分中草药浸提液在10倍稀释液条件下对小列当种子发芽的刺激或抑制作用最明显.研究发现,一些参试中草药蒸馏水和甲醇浸提液能够刺激或抑制小列当种子萌发,并且其对种子萌发的影响程度随浸提液浓度的变化而异,不同中草药最佳处理浓度不同,而且用甲醇浸提液可筛选出更多种具有化感作用的中草药.