Hydroxamic acid levels in Chilean and British wheat seedlings

Fifty-two cultivars of Chilean wheats were screened, using high pressure liquid chromatography, for hydroxamic acid levels at the seedling stage when maximal levels occur. DIMBOA levels ranged from 1.4 to 10.9 mmol/kg fr. wt and DIBOA levels ranged from 0 to 1.1 mmol/kg fr. wt. The family of wheat cultivars leading through breeding to the British cultivar Maris Freeman was examined. While DIMBOA and DIBOA levels in the ancestors ranged from 1.6 to 3.8 mmol/kg fr. wt and 0.16 to 0.34 mmol/kg fr. wt, respectively, DIMBOA and DIBOA levels in Maris Freeman were 2.39 and 0.24 mmol/kg fr. wt, respectively. The results are discussed in terms of the possibility of breeding for aphid resistance by making use of hydroxamic acids.     

Effect of content and distribution of hydroxamic acids in wheat on infestation by the aphid Schizaphis graminum

The content of hydroxamic acids in wheat plants shows substantial variations between different species and cultivars (1.0–6.3 mmol/kg fr. wt). It also varies with the age of the plant and the organ assayed. The maximum concentration is reached by the fourth day in epicotyls and roots. The amount in leaves at different plant ages is always higher in the younger leaves. Based on feeding and infestation experiments, it is proposed that the distribution of aphids on leaves of different ages is regulated by the hydroxamic acid content.     

A screen of worldwide wheat cultivars for hydroxamic acid levels and aphid antixenosis

In a screen of seedlings of a worldwide range of 47 cultivars of Triticum (mainly T. aestivum) the concentration of the hydroxamic acid DIMBOA ranged between 1 and 8 mmol/kg fresh wt. In a bioassay in which alatae of the aphid Sitobion avenae were released among replicated test seedlings, there were highly significant correlations between aphid ‘preference’ and DIMBOA levels in the seedlings. The value of these results in work leading to the production of aphid-resistant cultivars is discussed.     

Hydroxamic acids affecting barley yellow dwarf virus transmission by the aphid Rhopalosiphum padi

2,4-Dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA), a hydroxamic acid (Hx) occurring in wheat, was shown to deter feeding by the aphid Rhopalosiphum padi (L.), and to reduce BYDV transmission to the plant. Dual choice tests with wheat leaves showed the preferential settlement of aphids on leaves with lower levels of DIMBOA. Electric monitoring of aphid feeding behaviour showed that in seedlings with higher DIMBOA levels fewer aphids reached the phloem and they needed longer times to contact a phloem vessel than in those with lower levels. When aphids carrying BYDV were allowed to feed on wheat cultivars with different DIMBOA levels, fewer plants were infected with BYDV in the higher DIMBOA cultivars than in the lower ones. Preliminary field experiments showed a tendency for wheat cultivars with higher Hx levels to be more tolerant to infection by BYDV than lower Hx level ones.     

Distribution of hydroxamic acids in zea mays tissues

The hydroxamic acid content of leaves of cereals correlates well with resistance to aphids. In maize these compounds were absent from xylem exudates and guttation drops. Lateral veins of leaves of 7-day-old maize plants contained 8 mmol/kg fr. wt. while the entire leaf contained only 4.2 mmol/kg fr. wt. In leaves of 20-day-old plants, these amounts decreased by ca one-third. In mesocotyls, the cortex and central vascular cylinder contained 1.3 and 2.2 mmol/kg fr. wt, respectively. In 12-day-old wheat plants, the complete leaves and their veins contained 2.4 and 6.4 mmol/kg fr. wt respectively. Thus, the concentration of hydroxamic acid was always higher in the vascular bundles.     

The potential of hydroxamic acids in tetraploid and hexaploid wheat varieties as resistance factors against the bird‐cherry oat aphid,Rhopalosiphum padi

The potential for exploiting natural wheat resistance to control the cereal aphid Rhopalosiphum padi, the most important aphid pest of small grain cereals in the UK, was investigated as an alternative approach to the use of insecticides. The investigation focussed on a group of secondary metabolites, the hydroxamic acids or benzoxazinones, present naturally as glucosides, but which hydrolyse on tissue damage to give biologically active aglycones, e.g. 2,4‐dihydroxy‐7‐methoxy‐1,4‐benzoxazin‐3‐one (DIMBOA) which are associated with natural plant defence. These can be important for resistance against insects, fungi, bacteria and nematodes for a range of cultivated monocotyledonous plants and could ultimately be combined with other defence mechanisms to provide a general approach to cereal aphid control. Levels of hydroxamic acids, particularly DIMBOA‐glucoside, were determined in hexaploid (Triticum aestivum) and tetraploid (Triticum durum) wheat varieties and differences were found between species and varieties. The effect of feeding by R. padi on the level of hydroxamic acids in the leaf tissue was also investigated. Thus, after 24 h of aphid feeding, as an apparently localised hydrolytic defence reaction in the leaf, levels of DIMBOA‐glucoside decreased noticeably. When aphids were fed on sucrose solution containing low doses of DIMBOA there was a significant mortality compared to the sucrose control. However, the levels of and variation in hydroxamic acids in the wheat varieties investigated were insufficient for significant differences in aphid behaviour and development.     

Lack of C4 photosynthetic metabolism in ears of C3 cereals

There is continuing controversy over whether a degree of C₄ photosynthetic metabolism exists in ears of C₃ cereals. In this context, CO₂ exchange and the initial products of photosynthesis were examined in flag leaf blades and various ear parts of two durum wheat (Triticum durum Desf.) and two six-rowed barley (Hordeum vulgare L.) cultivars. Three weeks after anthesis, the CO₂ compensation concentration at 210 mmol mol^?1 O₂ in durum wheat and barley ear parts was similar to or greater than that in flag leaves. The O₂ dependence of the CO₂ compensation concentration in durum wheat ear parts, as well as in the flag leaf blade, was linear, as expected for C₃ photosynthesis. In a complementary experiment, intact and attached ears and flag leaf blades of barley and durum wheat were radio-labelled with ¹⁴CO₂ during a 10s pulse, and the initial products of fixation were studied in various parts of the ears (awns, glumes, inner bracts and grains) and in the flag leaf blade. All tissues assimilated CO₂ mainly by the Calvin (C₃) cycle, with little fixation of ¹⁴CO₂ into the C₄ acids malate and aspartate (about 10% or less). These collective data support the conclusion that in the ear parts of these C₃ cereals C₄ photosynthetic metabolism is nil.     

Hydroxamic acids in Secale cereale L. and the relationship with their antifeedant and allelopathic properties

Contents of the hydroxamic acids 2,4-dihydroxy-1,4-benzoxazin-3-one (DIBOA), and 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA) in leaves and roots of 14 cultivars of rye, Secale cereale L., were determined. Dynamics of accumulation in three cultivars were evaluated. DIBOA was the main cyclic hydroxamic acid in leaves but the contents differed significantly between the cultivars. Both DIBOA and DIMBOA were present in the roots. Maximum concentration of DIBOA in leaves and DIMBOA in roots was reached between 48-54 h and 54-72 h after germination, respectively. Antifeedant activity of DIBOA towards the aphid Rhopalosiphum padi and the feeding behavior were studied by electronic recording in barley leaves treated with different contents of DIBOA. The deleterious activity of DIBOA could arise by starvation and/or a toxic effect. Additionally, allelopathic potential of pure DIBOA and aqueous extracts of leaves and roots of rye (Tetra-Baer) on the germination of lettuce (Lactuca sativa) and rye (Tetra-Baer) seeds was evaluated. A high percentage of germination inhibition of pure DIBOA and the extracts of leaves and roots was observed. The activity is in agreement with the contents of hydroxamic acids in the plants. The substrates had no allelopathic effect on rye seeds.     

Occurrence and characterization of a UDP-glucose:hydroxamic acid glucosyltransferase isolated from wheat (Triticum aestivum) seedlings

Cyclic hydroxamic acid glucosides are present at high concentrations immediately after germination in wheat (Triticum aestivum L.). Changes in the activity of UDP-Glucose:cyclic hydroxamic acid glucosyltransferase (EC 2.4.1.-) in wheat were investigated using the cyclic hydroxamic acids 2.4-dihydroxy-1,4-benzoxazin-3-one (DIBOA) and its 7-methoxy derivative (DIMBOA) as sugar acceptors. Glucosyltransferase activity on both substrates was detected in dry seeds, with activity increasing after imbibition, peaking in shoots and roots 36-48 hours after imbibition and decreasing thereafter. The transience of glucosyltransferase activity was concurrent with the transient occurrence of the hydroxamic acid glucosides [Nakagawa E., Amano T., Hirai N., and Iwamura H. (1995) Phytochemistry 38, 1349-1354], suggesting that glucosyltransferases regulate the accumulation of hydroxamic acid glucosides in wheat seedlings. Two peaks in activity of UDP-Glucose:DIMBOA glucosyltransferase were detected using a Mono Q column, indicating the presence of at least two isozymes of this glucosyltransferase. The enzyme in the major peak was purified about 1500-fold and shown to be in a monomeric form with a molecular mass of 47 or 49 kDa. The enzyme reacted strongly with DIMBOA, less so with DIBOA. The enzyme of the minor peak on the Mono Q chromatogram, which was also a monomeric enzyme with a molecular mass of 47 kDa, showed similar substrate specificity to that of the major peak enzyme.     

Contents of 1,4-benzoxazin-3-ones and 2-benzoxazolinone from Stenandrium dulce (Nees)

Secondary metabolites, DIBOA, HBOA, 7-OH-HBOA and BOA, were isolated and quantified from S. dulce (Nees), a native species in Chile belonging to the Acanthaceae family. The highest DIBOA and HBOA contents were determined in leaves (9.25 mmol kg(-1) fr. wt) and root (6.81 mmol kg(-1) fr. wt), respectively. Aglycones, 7-OH-HBOA and HBOA, were isolated together from root extracts of Acanthaceae species. Both, HBOA and 7-OH-HBOA should be direct precursors in the biosynthesis of DIBOA and DIMBOA, respectively.     

Variation in Concentration of Ribulose-1, 5-bisphosphate Carboxylase in Leaves of Barley, Wheat and Hybrids of Crested Wheatgrasses

Amounts of the enzyme ribulose-1,5-bisphosphate carboxylasewere estimated in seedling leaves of barley (Hordewn vulgareL.) and flag leaves of wheat (Triticum aestitum L.) by radialimmuno diffusion. A fourfold variation among barley varietiesfor amount of RuBPCase at the seedling stage was observed (c.3.5–15mg g^–1 fr. wt). Range in variation for amountof flag leaf RuBPCase among wheat varieties was 6-09-9.39 mgRuBPCase g^–1 fr. wt. F₁ hybrids from interspecific andintergeneric crosses of crested wheatgrasses (Agropyron andElymus spp.) and their amphidiploid analogues were comparedfor amount of RuBPCase in the most recent fully expanded leavesharvested before seed set. Amount of enzyme varied from 3.4to 77.6 mg g^–1 fr. wt among the hybrids. No effect chromosomenumber on enzyme concentration was observed among 13 hybridsand their amphidiploid counterparts. Key words: RuBPCase, wheatgrasses     

不同环境条件下小麦氮代谢关键酶活性及籽粒品质

研究了两种环境条件下3个不同蛋白含量小麦品种的氮代谢关键酶活性及籽粒品质的差异.结果表明,龙口试验点的小麦旗叶硝酸还原酶(NR)、谷胺酰氨合成酶(GS)和籽粒谷胺酰氨合成酶活性均显著高于泰安试验点,3个品种间的酶活性顺序均为：济麦20＞优麦3号＞PH971942.优质强筋小麦品种的籽粒综合品质性状在龙口试验点的表现优于泰安试验点. 灌浆期环境因素与小麦籽粒品质和酶活性存在显著的相关性,灌浆期间的较高气温、适当干旱和寡照环境有利于提高小麦籽粒品质.龙口试验点的中、强筋小麦品种和泰安试验点的中筋小麦品种蛋白质含量与旗叶NR和GS活性均达显著正相关.小麦品种用途不同,对环境条件的要求不同,适宜的环境条件提高了氮代谢关键酶的活性,利于改善小麦品质.     

玉米化感物质异羟肟酸的研究进展

介绍了异羟肟酸在玉米植株中的分布和玉米根系分泌物中异羟肟酸的分析方法．丁布(DIM-BOA)是玉米植株中含量最大的异羟肟酸．不同玉米品种之间异羟肟酸含量的差异很大．种子不含异羟肟酸;但萌发后其含量迅速增加,在萌芽几天后的幼苗植株其含量达最大值,随后逐渐下降;在玉米生长发育的不同时期,幼嫩叶片内异羟肟酸含量始终较高;地上部分异羟肟酸的浓度高于根系．植株异羟肟酸的浓度受生长环境条件影响显著,在紫外辐射、黑暗条件或水分胁迫下其含量明显增加．在各种禾谷类作物中,玉米根系分泌物内含异羟肟酸较高;铁的存在能显著增加玉米根系分泌物中异羟肟酸的含量．     

Hydroxamic Acid glucosyltransferases from maize seedlings

Hydroxamic acids occur in several forms in maize (Zea mays L.) with 2,4-dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one (DIMBOA) being the predominant form and others including 2,4-dihydroxy-1,4-benzoxazin-3-one (DIBOA) being found at lower concentrations. Two enzymes capable of glucosylating hydroxamic acids were identified in maize protein extracts and partially purified and characterized. The total enzyme activity per seedling increased during the first 4 days of germination and was concurrent with the accumulation of DIMBOA. Purification of the enzymes by ammonium sulfate precipitation followed by Sephadex G-200 and Q-Sepharose gel chromatography resulted in a 13-fold increase in specific activity. The enzymes are initially separated into two peaks (peak 1 and peak 2) of activity by Q-Sepharose gel chromatography. The peak 1 glucosyltransferase had 3.6% of the DIMBOA glucosylating activity when DIBOA was used as substrate, whereas this percentage increased to 57% for the peak 2 enzyme. The enzyme in peak 2 has a K_m of 174 micromolar for DIMBOA and a K_m of 638 micromolar for DIBOA; the enzyme in peak 1 has a K_m of 217 micromolar for DIMBOA and its activity on DIBOA was too low to determine a K_m. The identification of two glucosyltransferases capable of glucosylating hydroxamic acids in vitro serves as an initial step in the characterization of the enzymes involved in production of hydroxamic acids in maize.     

Effects of DIMBOA levels in wheat on the susceptibility of the grain aphid (Sitobion avenue) to deltamethrin

The effects of DIMBOA in cultivars of wheat on aphid antibiosis and tolerance to the insecticide deltamethrin were investigated for the aphid Sitobion auenae. Over 48 hours the mean relative growth rate differed significantly between nymphs of S. auenae reared on wheat cultivars containing different levels of DIMBOA. Nymphs on the resistant cv. Altar showed a significantly greater susceptibility to deltamethrin, with nearly a three-fold increase in the relative toxicity of deltamethrin, compared with the susceptible cv. Dollarbird. The LC₅₀, adjusted for weight, was reduced by 91% for nymphs reared on the high DIMBOA cultivar. The potential for the enhancement of predation under a reduced pesticide régime is discussed in relation to this research.     

Involvement of Epicatechin in Cultivar Susceptibility of Avocado Fruits to Colletotrichum gloeosporioides after Harvest1

Avocado cultivars were defined as susceptible and resistant to Colletotrichum gloeosporioides depending upon the length of the incubation period of the disease after fruit softening. In the susceptible cultivars Fuerte, Horshim, Vurtz, Rincon, and Benik, epicatechin concentration of the peel decreased to 60-130 μg.g^?1, fr. wt. at fruit softening and symptoms appeared on the same or one day later. In the resistant cultivars Hass, Nabal, Netaim and Pinkerton, epicatechin concentration was still 632–1740 μg.g^?1 fr. wt. when fruit softening and symptoms appeared only 4-10 days later. When susceptible Fuerte fruits became soft the concentration of the antifungal compound 1-acetoxy-2-hydroxy-4-oxo-heneicosa-12,15 diene, had decreased to 120 μg.g^?1 fr. wt. and symptoms appeared. In resistant Hass fruits, the antifungal diene was still 238 μg.g^?1 fr. wt. at fruit softening; and it had further decreased to 159 μg.g^?1 fr. wt. when symptoms appeared, four days later. A modified atmosphere and 0.2 M CaCl₂ infiltration both delayed softening of Fuerte fruits; but symptom appearance on these fruits was related to diene decrease and not to fruit softening. The results are discussed in relation to the hypothesis that the susceptibility of avocado cultivars to post-harvest decay by C. gloeosporioides is related to the degradation of the antifungal diene, catalyzed by avocado lipoxygenase, the activity of which is regulated by the decline of its inhibitor epicatechin.     

Genotype and fungicide effects on late-season root growth of winter wheat

The aim of this work was to investigate differences among genotypes in post-anthesis root growth and distribution of modern UK winter wheat cultivars, and the effects of fungicide applications. Post-anthesis root growth of up to six cultivars of winter wheat (Triticum aestivum L.), given either one or three applications of fungicide, was studied in field experiments during two seasons. Total root mass remained unchanged between GS63 (anthesis) and GS85, but root length increased significantly from 14.7 to 31.4 km m⁻² in one season. Overall, there was no evidence for a decline in either root mass or length during grain filling. Root mass as a proportion of total plant mass was about 0.05 at GS85. There were significant differences among cultivars in root length and mass especially below 30 cm. Malacca had the smallest root length and Savannah the largest, and Shamrock had a significantly larger root system below 40 cm in both seasons. Fungicide applied at ear emergence had no significant effect on root mass in either season but increased root length (P<0.01) in the more disease-prone season. By maintaining a green canopy for longer, fungicide applied at flag leaf emergence may have resulted in delayed senescence of the root system and contributed to the post-anthesis maintenance of root mass and length. Section Editor: R. W. Bell     

Comparison of DIMBOA concentrations among wheat isolines and corresponding plant introduction lines

The concentration of a hydroxamic acid, also known as DIMBOA (2,4-dihydroxy-7-methoxy-1, 4-benzoxazin-3-one), in 6-d old wheat seedlings was examined using reverse-phase high performance liquid chromatography (HPLC). Wheat plant introduction (PI) lines PI 137739 (Dn1 gene), PI 262660 (Dn2 gene), and PI 294994 (Dn5 gene), the corresponding near-isogenic lines`Betta'-Dn1, Betta-Dn2 and Betta-Dn5, and susceptible Betta wheat were used in the study. The Dn2 gene conferring Russian wheat aphid, Diuraphis noxia (Mordvilko) (Hemiptera: Aphididae), tolerance was not related to DIMBOA concentration in wheat. Of the lines with Dn1 and Dn5 genes that confer antibiosis to D. noxia, only lines with the Dn5 gene showed increased DIMBOA accumulation. However, the Dn5 and the DIMBOA biosynthesis genes are not located in the same chromosome group. Possible relationship between the Dn5 gene and DIMBOA accumulation was discussed. This study indicates that DIMBOA concentration does not completely explain D. noxia resistance in the wheat lines examined and a comprehensive examination of other allelochemicals (e.g., phenolics) is necessary.     

Effect of salinity on phosphate accumulation and injury in soybean

Certain soybean [Glycine max (L.) Merr.] cultivars that are grown in saline nutrient cultures are killed when the inorganic phosphate (Pi) concentration in the substrate exceeds 0.10 mM. To determine the role of Na and Cl on this adverse salinity×Pi interaction, four cultivars, Clark, Clark 63, Lee, and Lee 74 were grown in the greenhouse in nutrient solutions salinized with 1) Cl and NO₃ salts to produce treatments with variable amounts of Cl or 2) with NaCl or KCl and CaCl₂ to obtain treatments with and without Na. At an osmotic potential of ?0.34 MPa, all salts enhanced Pi uptake and accumulation in the tissue of plants grown in ≧0.12 mM substrate Pi. Leaf Cl concentration was linearly related (r²≥0.9) to the mole fraction (mf) of Cl in the substrate, therefore excess substrate NO₃ did not greatly influence leaf Cl accumulation. Foliar injury was only observed on plants grown in saline solutions at high Pi (≥0.12 mM) and was not alleviated when KCl replaced NaCl in the substrate. This indicates that Na did not play a direct role in the salinity×Pi interaction. However, as the mf of Cl increased, severity of injury increased. The severity of injury, and its symptoms, were dependent upon leaf P and Cl concentration. Plants died when Cl and P in their leaves exceeded 800 and 600 mmol kg^?1 dry wt, respectively (e.g., Clark 63 grown at mf of Cl=1). The necrotic leaves were beige in color. Leaves that contained P in excess of 600 mmol kg^?1 dry wt and Cl between 150–200 mmol kg^?1 dry wt, were severely injured and reddish-brown in color (e.g., Clark 63 at mf of Cl=1/4 and Lee 74 Pi grown at mf of Cl=1). When leaf Cl was below 150 mmol kg^?1 dry wt, development of reddish-brown coloration in the leaves was sporadic. The adverse salinity×Pi interaction observed on these soybean variaties, therefore, was caused by a synergistic interaction between P and Cl in the leaves.     

Effects of Hydroxamic Acids Isolated from Gramineae on Adenosine 5'-triphosphate Synthesis in Chloroplasts

Two hydroxamic acids isolated from maize extracts, 2,4-dihydroxy-7-methoxy-1,4-(2H)-benzoxazin-3(4H)-one (DIMBOA) and the 2-O-beta-d-glucopyranoside of DIMBOA, inhibit photophosphorylation by spinach chloroplasts. Both cyclic and noncyclic photophosphorylations were inhibited to the same extent. The concentrations producing 50% inhibition for DIMBOA and its glucoside were about 1 and 4 millimolar, respectively. These compounds inhibit coupled electron transport but do not affect basal or uncoupled electron transport. Both acids inhibit the ATPase activities of membrane-bound coupling factor 1 (CF(1)) and of purified CF(1). On the basis of these results, it is concluded that DIMBOA and its glucoside act as energy transfer inhibitors of photophosphorylation.