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Relationship between SO2 dose and growth of the pea aphid, Acyrthosiphon pisum, on peas.

A series of experiments was carried out in controlled environment cabinets to investigate the effects of SO(2) dose on the mean relative growth rate (MRGR) of pea aphids, Acyrthosiphon pisum, feeding on the pea plant, Pisum sativum. There was a significant linear increase in the MRGR of aphids feeding on SO(2)-fumigated plants, relative to control aphids feeding on plants in charcoal-filtered air. The increase in MRGR reached a maximum of 11% at SO(2) concentrations between 90 and 110 nl litre(-1). MRGR declined at higher SO(2) concentrations until above 220 nl litre(-1) it was below that of the controls. The dose-response curve is discussed in relation to reported changes in the nitrogen metabolism of plants subject to air pollution, the response of aphids to host plant nitrogen and possible toxic effects of high concentrations of SO(2) on the aphid.

Journal Article↗

Toxicity to the pea aphid Acyrthosiphon pisum of anti-chymotrypsin isoforms and fragments of Bowman-Birk protease inhibitors from pea seeds.

Aphids feed on a protein-poor diet and are insensitive to several serine protease inhibitors. However, among the Bowman-Birk family of plant trypsin inhibitors (BBI), some members display significant toxicity to the pea aphid Acyrthosiphon pisum. A BBI isoform purified from pea seeds (PsTI-2) displays an IC50 of 41 microM and a LC50 of 48 microM at 7 days. Our data show that the chymotrypsin-directed active site from these bifunctional inhibitors is responsible for this activity, and that artificial cyclic peptides bearing the Bowman-Birk anti-chymotrypsin head induce much greater toxicity and growth inhibition than their anti-trypsin counterparts. The toxic syndrome included a rapid behavioural response of aphids on diets containing the toxic peptides, with induced restlessness after only 1 h of exposure to the chymotrypsin inhibitor. Nevertheless, chymotrypsin activity was not detected in aphid guts, using two chromogenic chymotrypsin substrates, and the physiological target of the chymotrypsin inhibitor remains unknown. These data show for the first time that plant chymotrypsin inhibitors, still widely unexplored, may act as paradoxical toxicants to aphids and serve as defensive metabolites for phloem-feeding insects.

Amino Acid Sequence↗

The cDNA cloning of a pea (Pisum sativum) seed lipoxygenase. Sequence comparisons of the two major pea seed lipoxygenase isoforms.

Cloning and sequencing of two cDNAs from mRNA of maturing pea (Pisum sativum) seeds allowed the deduction of the complete amino acid sequence of a lipoxygenase polypeptide which is most similar to that of soya-bean lipoxygenase 2. The predicted Mr of this polypeptide is 97134, and its sequence permits comparisons between the lox2-type and the lox3-type lipoxygenase isoforms from pea and soya bean (Glycine max).

Amino Acid Sequence↗

Purification of Multiple Forms of Glutathione Reductase from Pea (Pisum sativum L.) Seedlings and Enzyme Levels in Ozone-Fumigated Pea Leaves.

Glutathione reductase was purified from pea seedlings using a procedure that included 2',5'-ADP Sepharose, fast protein liquid chromatography (FPLC)-anion exchange, and FPLC-hydrophobic interaction chromatography. The purified glutathione reductase was resolved into six isoforms by chromatofocusing. The isoform eluting with an isoelectric point of 4.9 accounted for 18% of the total activity. The five isoforms with isoelectric points between 4.1 and 4.8 accounted for 82% of the activity. Purified glutathione reductase from isolated, intact chloroplasts also resolved into six isoforms after chromatofocusing. The isoform eluting at pH 4.9 constituted a minor fraction of the total activity. By comparing the chromatofocusing profile of the seedling extract with that of the chloroplast extract, we inferred that the least acidic isoform was extraplastidic and that the five isoforms eluting from pH 4.1 to 4.8 were plastidic. Both the plastidic (five isoforms were pooled) and extraplastidic glutathione reductases had a native molecular mass of 114 kD. The plastidic glutathione reductase is a homodimer with a subunit molecular mass of 55 kD. Both glutathione reductases had optimum activity at pH 7.8. The K(m) for the oxidized form of glutathione (GSSG) was 56.0 and 33.8 mum for plastidic and extraplastidic glutathione reductase, respectively, at 25 degrees C. The K(m) for NADPH was 4.8 and 4.0 mum for plastidic and extraplastidic isoforms, respectively. Antiserum raised against the plastidic glutathione reductase recognized a 55-kD polypeptide from purified antigen on western blots. In addition to the 55-kD polypeptide, another 36-kD polypeptide appeared on western blots of leaf crude extracts and the purified extraplastidic isoform. The lower molecular mass polypeptide might represent GSSG-independent enzyme activity observed on activity-staining gels of crude extracts or a protein that has an epitope similar to that in glutathione reductase. Fumigation with 75 nL L(-1) ozone for 4 h on 2 consecutive days had no significant effect on glutathione reductase activity in peas (Pisum sativum L.). However, immunoblotting showed a greater level of glutathione reductase protein in extracts from ozone-fumigated plants compared with that in control plants at the time when the target concentration was first reached, approximately 40 min from the start of the fumigation, and 4 h on the first day of fumigation.

Journal Article↗

Pea mania: deranged behaviour in cattle grazing a pea crop (Pisum sativum var arvense).

Hypersensitivity to external stimuli, progressing in some animals to manic behaviour, occurred in a cattle herd that grazed a crop of field peas (Pisum sativum var arvense) in the pre-flowering stage. Haematological and biochemical analyses eliminated hypomagnesaemia and ketosis as diagnoses. Other than two steers euthanased due to injuries sustained during manic episodes, all affected animals survived, recovering over 3 days when moved to alternative pasture. No necropsies were conducted. No microbial pathogens or endophytes were found on or in the plants. A previously reported incident in Victoria in 1987 in cattle grazing peas appeared to be of a similar nature. Environmental factors leading to these incidents were not clearly identified.

Animal Husbandry↗

Efficient intergeneric fusion of pea (Pisum sativum L.) and grass pea (Lathyrus sativus L.) protoplasts.

Large numbers of viable protoplasts of pea (Pisum sativum) and grass pea (Lathyrus sativus) were efficiently and reproducibly obtained and, for the first time, fused. Different procedures for fusion were compared, based either on electrofusion (750, 1000, 1250 or 1500 V cm(-1)), or on the use of macro or micromethods with a polyethylene glycol (PEG 6000 or PEG 1540), or a glycine/high pH solution. Over 10% of viable heterokaryons were obtained, with PEG as the most efficient and reproducible agent for protoplast fusion (>20% of viable heterokaryons). Both the division of heterokaryons and the formation of small calluses were observed.

Cell Fusion↗

[Pea (Pisum sativum) genes, participating in symbiosis with nitrogen-fixing bacteria. III. Study of the structure of the ENOD12 early nodulin gene for various types of peas using the polymerase chain reaction (PCR)].

We have determined the length of early noduline gene ENOD12 in various varieties and lines of pea (Pisum sativum) using the polymerase chain reaction (PCR). It was demonstrated that promoter regions of ENOD12A and ENOD12B genes in line 2150 (Afghanistan) are longer than these in variety "Feltham first". The disparity is 14 bp. When studying these genes in 7 different lines and varieties of pea it was found that ENOD12A gene is more variable in size than the ENOD12B gene. We showed the possibility to analyze the heritage of ENOD12 gene's alleles by using the PCR method.

Alleles↗

[Study on heredity laws of keratin membrane and sugar content in semi-leafless vegetable pea and their utilization in pea breeding].

It was studied for the heredity laws of keratin membrane and sugar content in semi-leafless vegetable pea. The results showed that keratin membrane character is controlled by two genes. F1 displays large keratin membranes in all fresh pods; F2 has three phenotypes, i.e. large keratin membrane, small keratin membrane and no keratin membrane with the ratio of 9:6:1. Sugar content is quantitative character fitting normal distribution. 'Xucai 1', a new semi-leafless vegetable pea variety with high yield and quality was successfully bred according to the laws. Its fresh pods taste delicious. Its extremely developed tendrils can be sold as top grade Longxu vegetable. The tender stems and leaves are also high quality vegetable.

Breeding↗

Pea seed-borne mosaic virus seed transmission exploits novel symplastic pathways to infect the pea embryo and is, in part, dependent upon chance.

Seed transmission of pea seed-borne mosaic virus (PSbMV) depends upon symplastic transport of the virus from infected maternal cells to the embryo. Such transport pathways have not been identified in higher plants. To identify these pathways, we have studied the ultrastructure of the tissues and cells around the micropyle of young developing seeds and compared transmitted and nontransmitted virus isolates. A characteristic of PSbMV infection was the presence of cylindrical inclusions positioned over plasmodesmal openings. The presence of cylindrical inclusions on the testa-endosperm boundary wall, together with immunogold labelling for virus-specific products on the wall and in the endosperm, indicated that symplastic connections existed at this interface. Close examination of the endosperm-suspensor boundary at the base of the suspensor revealed discontinuities in the suspensor sheath wall as porelike structures, which the virus might pass through en route to the embryo. A nontransmitted PSbMV isolate was able to invade the maternal tissues of the developing seed but was excluded from the embryo, although it was detected at a low level in the endosperm. Since the endosperm did not support virus replication, it appeared that passive accumulation determined the amount, timing, and location of the virus relative to the base of the suspensor. Rarely, therefore, could the nontransmitted virus isolate reach the correct location in the endosperm at the correct time for embryo infection via the suspensor to occur.

Embryo, Mammalian↗

Transient expression of a pea MAP kinase gene induced by gibberellic acid and 6-benzyladenine in unpollinated pea ovaries.

PsMAPK3, a new MAP kinase cDNA, was cloned from ovaries of Pisum sativum L. Expression of PsMAPK3 is at low basal levels in unpollinated ovaries but it is rapidly induced by gibberellic acid (peak at 30 min) and 6-benzyladenine (peak at 45 min). Both treatments promoted the development of a parthenocarpic fruit. In situ hybridization localized PsMAPK3 mRNA in ovules. The transcript was additionally detected in the mesocarp when it is expanding in response to the treatments. These observations suggest that gibberellins and cytokinins regulate PsMAPK3 mRNA levels in pea ovary shortly after fruit set is induced.

Adenine↗

Appearance of Three Chloroplast Isoenzymes in Dark-grown Pea Plants and Pea Seeds.

Activity peaks characteristic of the chloroplastic Calvin cycle enzymes triose-phosphate isomerase, ribose 5-phosphate isomerase, and fructose 1,6-diphosphate aldolase are found in isoelectric focusing patterns of dark-grown pea (Pisum sativum) seedlings and seeds. Apparently, in this higher plant these three chloroplastic isoenzymes can be formed in the absence of light and of chloroplast formation.

Journal Article↗

Pea Xyloglucan and Cellulose : III. Metabolism during Lateral Expansion of Pea Epicotyl Cells.

Lateral expansion of the third internodes of pea epicotyls was evoked by treatment with either 2,4-dichlorophenoxyacetic acid (2,4-D) or ethylene gas. During growth, 2,4-D enhanced and ethylene inhibited the deposition of xyloglucan and cellulose in the cell wall, with the result that the wall framework (ghost) from ethylene-treated swollen tissue was much thinner than that from 2,4-D-treated. The level of activity of xyloglucan synthase, alkali-insoluble beta-glucan synthases, and endo-1,4-beta-glucanases were all enhanced by 2,4-D treatment but not by ethylene. Both 2,4-D and ethylene treatments led to increased osmotic potential in the swelling tissues. Accordingly, swelling after 2,4-D treatment was accompanied by xyloglucan degradation, concomitant with substantial net synthesis, but swollen tissue as a result of ethylene treatment was characterized by walls whose integrity was weakened by relatively low levels of newly deposited polysaccharides rather than by the degradation.

Journal Article↗

Mechanisms underlying systemic invasion of pea plants by pea enation mosaic virus.

Leaves of Pisum sativum L.cv. 'Perfected Wales' were inoculated with pea enation mosaic virus, at the 3rd and 4th nodes as early as size permitted. Systemic virus spread was then measured over a period of approximately 9 days by assaying the infectivity of 1-cm stem pieces above the 4th and below the 3rd node. The presence of virus antigen and double-stranded (ds) RNA in these stem pieces was measured by radioimmunoassay over the same period. Relative infectivity and amounts of dsRNA in infected tissue were lowest approximately 3 days after inoculation and sharply increased thereafter. A transient plasmodesmatal abnormality was observed 4-6 days after inoculation. The transient nature of the abnormality, which was preceded (24 h) by low infectivity and dsRNA concentrations, may indicate a plant defense reaction that is subsequently overcome. The rise in dsRNA concentration, starting the 4th day after inoculation, coincided with the increase of vesicular material (cytopathological structures) in the phloem of infected plants. Vesicular material was found in sieve elements that were mature at the time of inoculation. Their presence in this tissue can only be explained by transport of the vesicles from infected sites.

Cytopathogenic Effect, Viral↗