Striga gesnerioidesParasitising Cowpea: Development of Infection Structures and Mechanisms of Penetration

Striga gesnerioides(Scrophulariaceae) is an obligate hemiparasiticangiosperm, that infects the roots of cowpea (Vigna unguiculata).The development ofStrigaseedlings was characterized after transferto the surface of cowpea roots using cryo-scanning electronmicroscopy. Contact with the surface of the root caused distinctmorphological changes in theStrigaradicles. Radicle elongationceased 8 h after contact with host roots and radicle hairs developed.These hairs adhered to the surface of the host root. Early stagesof root penetration, i.e. prior to connection to the host vascularsystem were examined by light microscopy and transmission electronmicroscopy to determine the mechanisms by whichStrigainvadesroot tissue. Cowpea roots were penetrated byS. gesnerioidesradiclesafter 48 h. Host cells immediately surrounding the invadingStrigawerecompressed, but other host cells were not affected. Immunocytochemistry,using monoclonal antibodies JIM 5 and JIM 7, showed thatStrigaadvancedintercellularly through the root tissues without extensive removalof pectins at the site of penetration.Copyright 1998 Annalsof Botany Company Striga gesnerioides,Vigna unguiculata, mechanisms of penetration, host-parasite interactions, electron microscopy, immunocytochemistry.     

Some Observations on Root-hair Infection by Nodule Bacteria

The infection by nodule bacteria of the root hairs of Viciahirsuta and of twelve species of Trifolium described. The proportion of root hairs deformed by bacterial secretionsvaries between host species and form point to point along theroot but is unaffected by bacterial strain. Infection is morefrequent in regions of maximum curling. Infection threads differin their place of origin in the hair (apical or lateral), theirmode of development and detailed morphology; some of these differencesare characteristic of host species. The growth of infectionthreads is often arrested in the root hair. Least arrested growthoccurs on 2-week-old seedlings infected at about twenty sites.The proportion of lateral infections increases with seedlingage. Root-hair curling, infection-thread initiation and growth areassociated with the near presence of the host cell nucleus. The observational data support the hypothesis of formation ofthe infection thread by invagination of the root-hair wall. No infections were observed in the root hairs of nodulated plantsof Lotus hispida, Lotus angustissimus, and Anthyllis vulneraria.     

Rhizobium Attachment and Curling in Asparagus, Rice and Oat Plants

Observations by scanning electron microscopy revealed that rhizobiaattach to the surface of rice protoplasts with regenerated cellwalls, isolated mesophyll cells of asparagus, and root hairsof rice and oat seedlings. Those strains of rhizobia, namelyRhizobium leguminosarum biovar trifolii, Bradyrhizobium japonicumand Bradyrhizobium sp., attach to the cells of these monocotsin the same manner as they attach to the host dicots tested.Escherichia coli did not attach. These results suggest thatthe attachment of rhizobia is not a host-specific process. Whenoat seedlings were infected by R. l. trifolii, hair curlingwas observed. The interactions between monocot plants and rhizobiais discussed in this paper. (Received June 12, 1989; Accepted November 9, 1989)     

Non-geotropic Growth Curvatures in the Striga Radicle

When seedlings of the root parasite Striga asiatica Benth. (=S. lutea Lour.) are grown in0 the absence of the host the radiclegrows to a length of about 4 mm. On emergence the radicle showsa strong curvature which is brought about by a failure of cellextension on one side. The inhibited cells generally remainun-extended throughout the life of the seedling. Neither thesodium salt of indole-3-acetic acid nor Inhibitor-ß(which is present in the seeds), induced permanent inhibitionsto the radicle cells when applied externally, as did the endogenouscurvature factor. 2: 3: 5-tri-iodobenzoic acid reduced curvatureby promoting extension of the normally inhibited cells.     

Analysis of In Vivo Protein Synthesis and Histological Studies of Haustorial Formation in Root Cultures of Witch weed (Striga asiatica L Kuntze)

We recently described an in vitro approach that uses root culturesto study haustorial formation in Striga asiatica. Previous studieshave used haustoria formed on intact radicles of Striga seedlings.In vitro cultured roots formed haustoria that appeared morphologicallysimilar to those formed by Striga radicles, but were 5–10-foldlarger. In this study, we provide biochemical and histologicalevidence to support further the similarity of root culture haustoriato haustoria formed on radicles of seedlings. We examined invivo protein synthesis during haustorial development on rootcultures and radicles by 2-D PAGE. Four proteins increased inabundance in both root cultures and radicles after 6 h of haustorialinduction. All four proteins appeared transiently in root culturesand radicles, being more abundant at 6 h, and less abundantafter 24 h of haustorial induction. Only three of the four haustorial-specificproteins were more abundant in root cultures after 2 h of haustorialinduction; all four had decreased in abundance after 12 h ofhaustorial induction. Using light microscopic analysis we comparedthe ontogeny of root culture haustoria to that of haustoriaon radicles. These studies revealed that root culture haustoriaundergo developmental changes similar to those reported forradicle haustoria such as early expansion of cortical cells,the emergence of haustorial hairs from epidermal cells, andthe development of densely staining cells at the haustorialapex. In addition, these changes occurred within a similar time-frameand sequence in root culture and radicle haustoria. Finally,root culture haustoria were found to be capable of attachingto sorghum host roots. Key words: Striga asiatica L., Kuntze, haustoria, root cultures, proteins, histology, 2D-PAGE     

Santalum album: Kernel Composition, Morphological and Nutrient Characteristics of Pre-parasitic Seedlings under Various Nutrient Regimes

Omission of all nutrients or individual minerals (N, K, P, S,Ca) from the growth medium of pre-parasiticSantalum album seedlingsproduced significant morphological effects. Compared with fullyfertilized seedlings, shoot and root length, leaf area, leaflength and width, leaf number, internode length, shoot length/rootlength ratio, chlorophyll levels, fresh mass and dry mass wereall reduced; leaf thickness increased and stomatal numbers,petiole lengths, root lengths and haustorial numbers showedvariable but significant responses. Mineral-specific deficiencysymptoms were evident in all omission treatments after 3 months.Treatments in which N, K or all minerals were omitted had thegreatest dwarfing effect and increase in height and leaf numberceased after 3 months: only seedlings with no added nutrients(dependent on kernel nutrition) were healthy. The mineral contentof no-nutrient seedlings approximated that in kernels. The distributionof minerals between plant parts for each treatment is given.K concentrations and K/Ca ratios were relatively high in leavesof seedlings which received treatments containing K, especiallywhen Ca was absent. A high K/Ca ratio may be inherent inS. albumseedlings rather than acquired after subsequent host connection.All fully fertilized seedlings developed haustoria within 3months suggesting readiness for early host attachment. Seedlingsin no-nutrient, no-N, no-K or no-P treatments had fewer haustoria. Santalum album ; Indian sandalwood; sandalwood kernels; sandalwood seedlings; nutrient omission; healthy characteristics; deficiency characteristics     

外生菌根真菌对重金属铜镉污染土壤中油松生长和元素积累分布的影响

该实验选取处于生长期的油松(Pinus tabulaeformis)幼苗,研究土壤Cu、Cd胁迫条件下,美味牛肝菌(Boletus edulis)单独接种、红绒盖牛肝菌(Xerocomus chrysenteron)与美味牛肝菌混合接种处理,对油松幼苗的生长和重金属积累分布状况的影响,探讨不同接种对油松抗性的影响。研究发现,菌根接种不仅促进寄主油松的生长发育和生物量积累,而且显著降低油松体内的重金属积累浓度,减少重金属由根部向植物茎叶部分的转运。与单一接种相比,混合接种可以更加有效地缓解重金属对寄主的生物毒性,减少土壤中重金属元素向油松体内的转运。这种优势在高浓度的重金属胁迫环境下尤为明显。该实验中,在3 mg·kg^-1 Cd胁迫下,混合菌根油松的茎叶和根部Cd浓度仅为未接种对照的59.1%和70.7%,比单一菌根降低了11.3%和18.1%,而混合菌根植物的茎叶和根部生物量干重则分别为未接种对照的1.14和1.20倍,单一菌根为未接种对照的1.18和1.17倍。在400 mg·kg^-1Cu胁迫下,混合菌根植物茎叶和根部的干重分别是未接种植株的1.01和1.09倍,而混合菌根植物茎叶和根部的Cu浓度仅为未接种植株的61.8%和79.6%,比Boletus edulis菌根植物的Cu积累浓度下降了0.7%和3.8%。     

Mineral Nutrition of Sandalwood (Santalum spicatum)

Acacia acuminata is a preferred host of the root hemiparasitictree, Santalum spicatum (sandalwood). Comparison between nutrientcontent of adult trees of sandalwood and results for an earlierstudy of the mistletoe, Amyema preissii, on the same host species,A. acuminata, showed similar high levels of K and Na and lowlevels of Zn in both parasites compared with the host plants.Differences in K, Ca, N and Cu levels between parasitized anduninfected Acacias imply that the host plant contributes tothe nutrition of sandalwood. The high K/Ca ratio in sandalwoodconfirms that K uptake in preference to Ca is a general featureof all categories of angiosperm parasites. Patterns of distribution of nutrients between various partsof sandalwood and A. acuminata depend on the type of nutrient,but levels are usually highest in leaves of both species andthe haustoria. Although K, Ca and Na are much lower in the kernelsthan in vegetative parts of the parasite, only seedlings withoutsupplementary Ca in a nutrient omission experiment failed togrow at all in the absence of hosts. Growth is not dependenton the level of K in the unattached plants but other evidenceindicates it may have a role in water uptake in the attachedplant. Calcium supply has a marked effect on internal Ca levelsand growth of unattached plants. Compared with field plants,levels of Ca, and to a lesser extent Zn, were much higher inplants of the Ca/K treatment that produced greatest growth over34 weeks. Haustorial formation is enhanced by the presence of A. acuminataroots. However, competition for nutrients, especially Ca, fromco-planted A. acuminata seedlings results in suppression ofgrowth of young sandalwood compared with their growth in theabsence of the host species. Key words: Mineral nutrients, Santalum spicatum, Acacia acuminata, hemiparasites, K/Ca nutrition, seeds     

Root Growth, Secondary Root Formation and Root Gravitropism in Carotenoid-deficient Seedlings of Zea mays L.

The effect of ABA on root growth, secondary-root formation androot gravitropism in seedlings of Zea mays was investigatedby using Fluridone-treated seedlings and a viviparous mutant,both of which lack carotenoids and ABA. Primary roots of seedlingsgrown in the presence of Fluridone grew significantly slowerthan those of control (i.e. untreated) roots. Elongation ofFluridone-treated roots was inhibited significantly by the exogenousapplication of 1 mM ABA. Exogenous application of 1 µMand 1 nM ABA had either no effect or only a slight stimulatoryeffect on root elongation, depending on the method of application.The absence of ABA in Fluridone-treated plants was not an importantfactor in secondary-root formation in seedlings less than 9–10d old. However, ABA may suppress secondary-root formation inolder seedlings, since 11-d-old control seedlings had significantlyfewer secondary roots than Fluridone-treated seedlings. Rootsof Fluridone-treated and control seedlings were graviresponsive.Similar data were obtained for vp-9 mutants of Z. mays, whichare phenotypically identical to Fluridone-treated seedlings.These results indicate that ABA is necessary for neither secondary-rootformation nor for positive gravitropism by primary roots. Zea mays, gravitropism, carotenoid-deficient, Fluridone, root growth, vp-9 mutant     

Factors Influencing Adventitious Root Production in Cuttings of Griselinia littoralis and Griselinia lucida

Griselinia littoralis roots quickly and vigorously from cuttingsof seedlings and mature plants and also forms roots on detachedleaves. Cuttings root in the dark but leaves must be present.In contrast G. lucida roots vigorously only from cuttings takenfrom seedlings. Light is essential for root formation on oldermaterial. Detached leaves will not root. Although callus formationat the cut base commonly occurs in both species it is not directlyassociated with root formation. Griselinia littoralis Raoul Choix, Griselinia lucida Forst. f. Prodr., adventitious roots, stem cuttings, leaf cuttings, woody plants     

Inoculation with Azospirillum lipoferum Affects Growth and Gibberellin Status of Corn Seedling Roots

Maize (Zea mays L., hybrid Cargill 147) seedlings, grown inaseptic conditions, were inoculated with three strains of Azospirillumlipoferum (Al op 33, Al iaa 320, and ATCC 29708) or culturedin different concentrations of indol-3-acetic acid (IAA) orgibberellin A₃ (GA₃). After 48 h, root length, root surfacearea, root dry weight, and shoot dry weight were measured inall treatments. Gibberellin content was evaluated in selectedroots of control and Azospirillum inoculated seedlings by gaschromatography-mass spectrometry-selected ion monitoring withthe use of deuterated gibberellins as internal standards. Thethree strains of A. lipoferum, IAA (2 ng ml^–1), and GA₃(40 to 400 pg ml^–1) significantly enhanced root growth.Improvement of root hair growth and density was obtained mainlywith A. lipoferum strain Al op 33 and GA₃ 40 pg ml^–1.GA₃ was identified by gas chromatography-mass spectrometry (byboth, full scan and selected ion monitoring) in a free acidfraction from roots of the seedlings inoculated with A. lipoferum.In the roots of the non inoculated seedlings GA₃ was found afterhydrolysis of a fraction expected to contain glucosyl conjugates. (Received April 26, 1993; Accepted September 27, 1993)     

Biological and Cultural Characteristics of the Effective Frankia Strain HFPCcl3 (Actinomycetales) from Casuarina cunninghamiana (Casuarinaceae)

From homogenates prepared from surface-sterilized nodules ofseedlings of Casuarina cunninghamiana grown aeroponically, astrain of Frankia designated HFPCc13 was isolated and has beengrown in pure filamentous culture in a defined synthetic nutrientmedium. Vesicle and sporangium formation can be induced by removalof combined nitrogen from the medium.Frankia strain HFPCc13nodulates young seedlings of C. cunninghamiana and C. equisetifoliawithin three weeks of inoculation with an optimum root mediumpH of 6–7 for nodulation and optimum temperature of 30–35^°C. The presence of combined nitrogen in the root mediuminhibits nodulation with NH₄⁺ more inhibitory than NO₃^–.Frankia HFPCc13 does not nodulate Allocasuarina species withinthe same family nor several other possible actinorhizal plantstested. Thus this strain is quite precise in its host specificity.The rate of acetylene reduction was greater in C. cunninghamianathan the closely related species C. equisetifolia. In neitherof these host species were vesicles observed to occur withinthe infected root nodules which had been demonstrated to beactively fixing dinitrogen. Root nodules were shown to be activein acetylene reduction over a range of O₂ concentration in thegaseous environment with an optimum at about 20 per cent O₂,the ambient P_O2 of the air. The mechanism(s) for oxygen protectionof nitrogenase within the filamentous form of Frankia withinthese nodules remains to be explained. Casuarina, Frankia, nodulation, nitrogen fixation     

The possible involvement of a spore agglutinating factor(s) in various plants in establishing host specificity by various strains of black rot fungus, Ceratocystis fimbriata

Extracts, possibly containing a factor(s) which agglutinatesspores of Ceratocystis fimbriata, were obtained from the sweetpotato, potato, taro, cucumber and kidney bean. The factor(s)seemed to be involved in the expression of host specificityby various strains of C. fimbriata. The agglutinating factor(s)from sweet potato root had a high molecular weight and was inactivatedby pronase or macerozyme treatment. (Received March 6, 1974; )     

Morphological Response of Witchweed (Striga asiatica) to In Vitro Culture

Excised sorghum root segments (5–10 mm in length) werecultured for 50 d in four different liquid media containingmineral salts, vitamins, amino acids, glucose, and IAA. Theroots were removed and the remaining medium was solidified withan equal volume of warm 1–6% water agar. Dry unconditionedor conditioned Striga asiatica seeds were transferred to themedium. Some of the seeds germinated and developed into parasitic-typeseedlings. These seedlings had haustoria, tubercles, dense roothairs, branched shoots, and multiple shoot-borne adventitiousroots. The plumule pole developed into a shoot, but the radiclepole displayed only rudimentary development. On the same media,but which had not previously been used to grow sorghum roots,the seedlings displayed a well-developed radicle-derived rootsystem, but the plumule did not grow. Shoots began to appearon the roots only after 35–50 d of culture. These seedlingshad no haustoria, no tubercles, few or scattered root hairs,no shoot-borne adventitious roots and few shoot branches, andappeared to be non-parasitic-type seedlings. Shoots grew ina medium supplemented with IAA and kinetin, but did not in amedium containing NAA plus IBA. On replacement of glucose andIAA with sucrose and 2,4-D, respectively, Striga seeds germinated,and the heart-shaped embryos dedifferentiated into calli. Thecalli have been maintained by subculturing for over 9 months.The results demonstrated that a host signal, in addition tothose for germination and haustorium formation, is requiredfor further development. Moreover, morphogenesis of culturedS. asiatica is influenced by exogenous growth regulators. Key words: Striga asiatica, parasitic weeds, haustoria, Sorghum bicolor, in vitro culture     

Anatomy, Growth and Survival of a Long-hypocotyl Mutant of Cucumis sativus Deficient in Phytochrome B

Plant dry matter (DM) partitioning, survival rates, stem anatomy,and stem water conductivity were investigated in wild-type (WT)and long-hypocotyl (lh) mutant seedlings of cucumber (Cucumissativus) grown as isolated individuals under natural radiation.The lh mutant is severely deficient in phytochrome B. Wild-typeseedlings accumulated more DM than lh seedlings over a 4-weekgrowth period in the glasshouse. Leaf and root DM were higherin the WT but stem DM was higher in the lh mutant. Stem DM perunit length was larger in WT than in lh mutant seedlings, evenwhen the two genotypes were compared at equal whole plant DM,which was achieved by growing the plants under different irradiance.In WT seedlings, the hypocotyl was shorter but thicker, withlarger average cell diameter than the lh mutant. In hypocotyltransverse sections the area occupied by load-bearing tissues(xylem and phloem fibres) and the number and diameter of xylemvessels were larger in WT than lh seedlings. Survival ratesof the lh mutant were normal in the glasshouse but very lowoutdoors due to hypocotyl fracture. The water conductivity ofhypocotyl sections was higher in WT than lh seedlings, but nosignificant differences in water conductivity were observedwhen the root remained attached to the hypocotyl. These resultssuggest (a) that compared to the WT, tall and slender lh plantsare more susceptible to mechanical stresses created by windimpact, and (b) that if the lh lesion affects the phyB geneonly, phytochrome B plays a role in the elicitation of anatomicaland morphological changes that specifically increase fitnessin open environments.Copyright 1994, 1999 Academic Press Cucumis sativus (cucumber), light phenotypes, phytochrome, photomorphogenesis, shade phenotypes, stem growth     

The Initiation of the Secondary Haustorium in Alectra vogelii Benth

Anatomical studies were carried out on initiation of the secondaryhaustorium in Alectra vogelii, a root parasite of leguminouscrops in Nigeria. In both the normal and self-haustorium, theformation of the haustorial initial on the parasite root soonafter initial contact between the host and parasite roots isfollowed by the penetration of the host root by the haustorium.Specialized penetrating cells (intrusive cells) at the haustorialfront prise apart and loosen the host root cortical cells, whichlater become digested. Through the same processes, a few ofthese columnar intrusive cells at the haustorial front piercethe endodermis to make contact with the xylem of the host root.Thereafter, a true conductive bridge consisting of short, isodiametric,reticulate vessel elements is established between the parasiteand host roots through the secondary haustorium. No pholem tissuewas observed in the connection. There is a close similaritybetween the mode of initiation of the secondary haustorium ofAlectra vogelii and that previously described for its primaryhaustorium. Alectra vogelii Benth, haustorium, self-haustorium, root parasite, hemiparasitism, Vigna unguiculata, Arachis hypogaea     

Haustoria of Cuscuta japonica, a Holoparasitic Flowering Plant, Are Induced by the Cooperative Effects of Far-Red Light and Tactile Stimuli

When seedlings of Cuscuta japonica were grown with Vigna radiata(the host plant) in a flower pot for 6 d under white light andthen irradiated with far-red or blue light (ca. 6 µmolphotons m^–2 s^–1), the seedlings parasitized V. radiata.However, no parasitism of the seedlings was observed under redor white light or in darkness. The parasitic behavior of seedlingsof C. japonica was observed even if an acrylic rod was usedas a substitute for the host plant. Upon incubation under far-redlight, the seedling twined tightly around the rod and developedhaustoria towards it. Haustoria also developed when apical andsubapical regions of seedlings were held between two glass platesthat were about 0.7 mm apart and were irradiated with far-redlight. However, no haustoria were induced by either the holdor irradiation alone. These results indicate that parasitismof Cuscuta japonica is controlled by the cooperative effectsof two physical signals, far-red light and appropriate tactilepressure. Our findings suggest that parasitism by the genusCuscuta involves a novel strategy. (Received April 10, 1996; Accepted August 21, 1996)     

Structure and Development of the Mature Secondary Haustorium in Alectra vogelii Benth

Anatomical investigations were carried out on the structureand development of the mature secondary haustorium in Alectravogelii growing on Arachis hypogaea or Vigna unguiculata. Followingthe formation of the young secondary haustorium, both the cambiumand pericycle of the host root directly opposite the young secondaryhaustorium are stimulated to divide and form new tissues andorgans including haustorial roots. Further proliferations ofthe host root pericycle and the haustorial cortex give riseto a large, tuberous and complex mature secondary haustoriumwithin which the tissues of the host and parasite remain inintimate contact forming a perfect graft union with a wide zoneof contact. Apart from the haustorial axial xylcm strand whichnormally connects the xylem of the parasite secondary root withthat of the host, direct xylary connections are also establishedbetween the axial xylem of the haustorium and the xylem of thehaustorial roots. The entire surface of the mature secondaryhaustorium of Alectrais covered with these haustorial rootsas was previously observed in its mature primary haustorium. Alectra vogelii Benth, secondary haustorium, haustorium, haustorial roots, root parasite, hemiparasitism, Arachis hypogaea, Vigna unguiculata     

Growth and Graviresponsiveness of Primary Roots of Zea mays Seedlings Deficient in Abscisic Acid and Gibberellic Acid

Moore, R. and Dickey, K. 1985. Growth and graviresponsivenessof primary roots of Zea mays seedlings deficient in abscisicacid and gibberellic acid.—J. exp. Bot. 36: 1793–1798. The objective of this research was to determine if gibberellicacid (GA) and/or abscisic acid (ABA) are necessary for graviresponsivenessby primary roots of Zea mays. To accomplish this objective wemeasured the growth and graviresponsiveness of primary rootsof seedlings in which the synthesis of ABA and GA was inhibitedcollectively and individually by genetic and chemical means.Roots of seedlings treated with Fluridone (an inhibitor of ABAbiosynthesis) and Ancymidol (an inhibitor of GA biosynthesis)were characterized by slower growth rates but not significantlydifferent gravicurvatures as compared to untreated controls.Gravicurvatures of primary roots of d-5 mutants (having undetectablelevels of GA) and vp-9 mutants (having undetectable levels ofABA) were not significantly different from those of wild-typeseedlings. Roots of seedlings in which the biosynthesis of ABAand GA was collectively inhibited were characterized by gravicurvaturesnot significantly different from those of controls. These results(1) indicate that drastic reductions in the amount of ABA andGA in Z. mays seedlings do not significantly alter root graviresponsiveness,(2) suggest that neither ABA nor GA is necessary for root gravicurvature,and (3) indicate that root gravicurvature is not necessarilyproportional to root elongation. Key words: Abscisic acid, Ancymidol, Fluridone, gibberellic acid, root gravitropism, Zea mays     

Hair Curling Induced in Heterologous Legumes and Monocots by Flavonoid-Treated Rhizobia

The curling of root hairs and the deformation response wereobserved when white clover was infected with homologous strainsof Rhizobium leguminosarum biovar trifolii 4S and 0403. In thecase of Rhizobium meliloti NZ and Rhizobium leguminosarum biovarviciae 128C53, however, curling was only induced when thesebacteria were pretreated with flavonoids: luteolin in the caseof R. meliloti and naringenin for R.I. viciae. The same resultswere obtained with oat, a monocotyledonous non-leguminous plant.The two flavonoids mentioned are secreted from the host plantsand induce the expression of genes for root hair curling (Hac)on Sym plasmid in homologous rhizobia, therefore, the curlingresponse in both white clover and oat appears to be correlatedwith the activation of the Hac genes. These results suggestthat a factor(s) that activates the Hac genes, such as 7,4'-dihydroxyflavonewhich is known as the factor required by R. I. trifolii, issecreted from the oat roots. (Received June 12, 1989; Accepted November 9, 1989)