Ethylene-promoted ascorbate peroxidase activity protects plants against hydrogen peroxide, ozone and paraquat

Abstract. In experiments where mung beans ( Vigna radiata L.) and peas ( Pisum sativum L.) have been pre-exposed to ethylene and afterwards treated with ozone, it has been shown that such ethylenepretreated plants may become more resistant to ozone. Further experiments with hydrogen peroxide (H₂O₂) and the herbicide paraquat suggest that this increased resistance against ozone depends on the stimulation of ascorbate peroxidase activity which provides cells with increased resistance against the formation of H₂O₂ which is also formed when plants are fumigated with ozone. These results explain why increased production of ethylene can be observed in plants exposed with ozone or other oxidative stress and clearly demonstrate that in plants, as well as animals, peroxidases protect cells against harmful concentrations of hydroperoxides.     

Temperature influence on cohort parameters and demographic characteristics of the two cowpea coreids Clavigralla tomentosicollis and C. shadabi

Age-specific life tables of two important pests of cowpea, Vigna unguiculata (L.) Walp., the pod sucking bugs Clavigralla tomentosicollis Stål and C. shadabi Dolling (Heteroptera: Coreidae), were obtained from observations carried out at different temperatures. A biophysical model was found satisfactory to describe the temperature-response of developmental and mortality rates of egg and nymphal stages, with a peak developmental rate around 34°C in both species. The variability in development times was small and the experimental data did not permit any conclusion with regard to the Erlang probability density function. Survival of eggs and nymphs remained high between 20° and 30°C for both species. At temperatures above 34°C, C. tomentosicollis survivorship and fecundity was higher than that of C. shadabi, which in turn laid more eggs at temperatures between 20° and 30°C. Maximum fecundity is estimated to be at 29°C for C. tomentosicollis (99 eggs/female) and 26°C for C. shadabi (261 eggs/female). At 30°C, the intrinsic rate of increase reached a maximum in both species, 0.152 per day for C. tomentosicollis and 0.145 per day for C. shadabi, and remained high for C. tomentosicollis until 36°C. C. tomentosicollis performed significantly better on pigeonpea, Cajanus cajan Millsp., than on cowpea at higher temperatures.     

Promoter regions of cysteine endopeptidase genes from legumes confer germination-specific expression in transgenic tobacco seeds

Cysteine endopeptidases, SH-EP from Vigna mungo and EP-C1 from Phaseolus vulgaris, act to degrade seed storage protein during seed germination. Using transgenic tobacco plants, expression of SH-EP and promoter activity of the EP-C1 gene were analyzed in transgenic tobacco plants. The promoters of the two genes in tobacco seeds showed germination-specific activation, although post-translational processing of SH-EP and regulatory regions of promoter of the gene for EP-C1 were found to differ between leguminous seeds and transgenic tobacco seeds.     

Isolation and cDNA cloning of an Em-like protein from mung bean (Vigna radiata) axes

Until now no 'early-methionine-labelled' (Em) proteins have been reported in the Fabaceae. To check whether a previously isolated low-molecular mass albumin from dry mung bean embryonic axes possibly corresponded to an Em-like protein, the protein was purified, sequenced and its cDNA clone isolated and characterized. N-terminal sequencing of cyanogen bromide cleavage products of the protein revealed homology with previously described Em-like proteins from other species. Analysis of cDNA clones encoding the mung bean Em protein revealed the presence of two classes of Em proteins and confirmed their homology to the previously characterized Em-like proteins. In vivo labelling and northern blot analysis further demonstrated that the mung bean protein is synthesized during early germination of the axes and that abscisic acid (ABA) extends its synthesis. It appears, therefore, that legumes also contain maturation-specific, ABA-responsive Em-like proteins.     

Impact of density of Aphis craccivora (Aphididae) on growth and yield of susceptible and resistant cowpea cultivars

Impact of initial density of cowpea aphid Aphis craccivora Koch (Aphididae) at infestation on the growth and yield of aphid-susceptible cowpea cultivar ICV-1 and aphid-resistant cultivar ICV-12, was investigated. Plants at the seedling, flowering and podding stages of development were infested with five aphid densities consisting of 0, 2, 5, 10 and. 20 aphids per plant and maintained for 22 days. Extended leaf heights of plants and aphid counts were recorded at 7, 12, 17 and 22 days after infestation. Two crop growth parameters (biomass duration and leaf area duration), and two plant yield parameters (number of pods per plant and number of seeds per pod) were recorded. Due to the occurrence of parthenogenesis and changes in population dynamics during infestations, aphid densities were converted into cumulative cowpea aphid-days, to facilitate data analyses and interpretation. ANOVA indicated that there was significant (P=s 0.05) difference in aphid-day accumulations between the two cultivars when infested at the seedling stage. Accumulations on cv. ICV-1 were greater than on cv. ICV-12. However, no such differences between the cultivars were detected when plants were infested at flowering and podding stages. Therefore, the seedling stage was used for comparisons of the impact of cowpea aphid-days on the growth and yield parameters of the two cultivars. At the 95% confidence intervals, ICV-12 plants were consistently taller than ICV-1 plants. Infested ICV-1 seedlings showed stunting and other growth deformities which were not observed on ICV-12 plants. Regression analyses revealed substantial reductions in the growth and yield parameters of ICV-1 relative to ICV-12. Overall, cowpea aphid-days provided a convenient and reliable method for studying the aphid population dynamics and the subsequent impact on plant growth and yield performance.     

Characterization of two cDNAs for novel drought-inducible genes in the highly drought-tolerant cowpea

Ten cDNAs for drought-inducible genes were isolated using differential screening of a cDNA library prepared from 10-hr dehydrated cowpea plants,Vigna unguiculata (S. Iuchi, K. Yamaguchi-Shinozaki, T. Urao, T. Terao, K. Shinozaki; Plant Cell Physiology, 1996 in press). Two of the cDNA clones, designated CPRD12 and CPRD46, were sequenced and characterized. The CPRD12 and CPRD46 cDNAs encode putative proteins related to nonmetallo-short-chain alcohol dehydrogenase (CPRD12) and chloroplastic lipoxygenase (CPRD46). Northern blot analysis revealed that these genes are induced by high-salinity stress and exogenous abscisic acid, but not by cold stress. The CPRD46 gene is also responsive to heat stress and methyl jasmonate and salicylic acid. Genomic Southern blot analysis suggested that CPRD12 constitutes a small gene family, but that CPRD46 is a single copy gene. We discuss the possible functions of these two CPRD gene products under drought stress.     

Molecular heterogeneity of Cowpea (Vigna unguiculata Fabaceae) seed storage proteins

81 wild forms and 110 cultivated cowpea,Vigna unguiculata, accessions from 21 countries of Africa were screened for variability in seed storage proteins. Total seed proteins, albumin and globulin fractions were investigated by means of sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and isoelectric focusing (IEF) of nonreduced and/or reduced samples in one- and two-dimensional procedures. The globulin fraction is heterogeneous in molecular weight and contains both legumin-like components and three to six nondisulfide-linked subunits. Three globulin subunits, with molecular weights 110, 76, and 41 kD were found to be composed of disulfide-linked polypeptides. In the nondisulfide-linked fraction, both cultivated and wild forms exhibited patterns of four types (A–D). This fraction contains polypeptide subunits of molecular weights 62, 56, and 52 kD for A type, 62, 56, 54, and 52 kD for B type, 62, 56, 52, and 50 kD for C type, and at least 62, 56, 54, 52, 50, and 49 kD for D type. These subunits present similar multiple charge forms but C and D types possess more basic specific 50 and 49 kD nondisulfide linked components. Major albumin fraction contains subunits of 94, 86, 32, and 24kD. No infraspecific variation was observed in albumin or legumin-like fractions. The discussion is focussed on the relations between genetic variability assessed by storage protein coding genes and phenotypic variability.     

Acid-induced changes in the in vivo wall-yielding properties of hypocotyl sections of Vigna unguiculata

Elongation growth of hypocotyl sections of Vigna unguiculata under xylem perfusion was significantly enhanced when acid was applied by acid-aerosol to an abraded hypocotyl surface in the air. The in vivo wall extensibility (φ) and the effective turgor (Pⁱ– Y), both of which were determined by the pressure-jump method, increased during acid-induced growth as observed in IAA-induced growth. The intracellular pressure (Pⁱ), however, decreased significantly at the beginning of acid-induced growth whereas Pⁱ scarcely changed in IAA-induced growth. This result indicates that protons increase the effective turgor by decreasing the yield threshold as IAA does. There seems to be no essential difference between proton and auxin in the effects on the in vivo mechanical properties of the surface cell wall.     

Selection and characterization of mannitol-tolerant callus lines of Vigna radiata (L.) Wilczek

An osmotically (mannitol) tolerant callus line of Vigna radiata (L.) Wilczek has been isolated from callus cultures grown on modified PC-L2 medium supplemented with increasing concentrations of mannitol. The tolerance was stable and retained after growth in the absence of mannitol selection for 2 months. The growth of the tolerant line, in the presence of mannitol (540 mol m^-3) was comparable to that of a sensitive callus line growing in the absence of mannitol. This line not only grew well on media containing up to 720 mol m^-3 mannitol, but also required 450 mol m^-3 mannitol for its optimal growth. Osmotically tolerant callus also showed increased tolerance to NaCl (0–250 mol m^-3) stress as compared to sensitive callus. Accumulation of Na⁺ was lower, and the level of K⁺ was more stable in osmotically tolerant than in sensitive calli, when both were exposed to salt. The free proline content of both tolerant and sensitive calli increased on media supplemented with mannitol or NaCl. However, the proline content of sensitive callus was higher than in tolerant callus in the presence of same concentrations of mannitol or NaCl.Abbreviations NAA -naphthaleneacetic acid - 2,4-d 2,4-dichlorophenoxyacetic acid - BAP 6-benzylaminopurine     

Variation at isozyme loci in wildVigna unguiculata (Fabaceae,Phaseoleae)

An electrophoretic comparison of variation at 37 presumptive isozyme gene loci was performed for 55 wildVigna unguiculata accessions. The analysis included seven subspecies, covering the whole range of variation available in wildVigna unguiculata. The results of the isoenzymatic study broadly confirmed the previous infraspecific classification inferred from morphological data. Although the southern taxa (e.g., subsp.tenuis, subsp.stenophylla) are as yet too poorly sampled to discuss in depth, wildVigna unguiculata can be divided into one mainly autogamous annual group (including subsp.pubescens and var.spontanea) and several widely divergent perennial subspecies.