J. E. Harper

Publications (5)23.75 Total impact

• Source

  Available from: plantphysiol.org

  Article: Shoot versus Root Signal Involvement in Nodulation and Vegetative Growth in Wild-Type and Hypernodulating Soybean Genotypes.

  C. Sheng, J. E. Harper
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  ABSTRACT: Grafting studies involving Williams 82 (normally nodulating) and NOD1-3 (hypernodulating) soybean (Glycine max [L.] Merr.) lines and Lablab purpureus were used to evaluate the effect of shoot and root on nodulation control and plant growth. A single- or double-wedge graft technique, with superimposed partial defoliation, was used to separate signal control from a photosynthate supply effect. Grafting of hypernodulated soybean shoots to roots of Williams 82 or L. purpureus resulted in increased nodule numbers. Grafting of two shoots to one root enhanced root growth in both soybean genotypes, whereas the nodule number was a function of shoot genotype but not of the photosynthetic area. In double-shoot, single-root-grafted plants, removing trifoliolate leaves from either Williams 82 or NOD1-3 shoots decreased root and shoot dry matter, attributable to decreased photosynthetic source. Concurrently, Williams 82 shoot defoliation increased the nodule number, whereas NOD1-3 shoot defoliation decreased the nodule number on both soybean and L. purpureus roots. It was concluded that (a) soybean leaves are the dominant site of autoregulatory signal production, which controls the nodule number; (b) soybean and L. purpureus have a common, translocatable, autoregulatory control signal; (c) seedling vegetative growth and nodule number are independently controlled; and (d) two signals, inhibitor and promoter, may be involved in controlling legume nodule numbers.
  Plant physiology 04/1997; 113(3):825-831. · 6.53 Impact Factor
• Article: Identification of cDNA clones corresponding to two inducible nitrate reductase genes in soybean: analysis in wild-type and nr1 mutant.

  S Wu, Q Lu, A L Kriz, J E Harper
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  ABSTRACT: Among higher plants, soybean is unique in that biochemically it has been characterized as having two constitutive nitrate reductase (cNR) isoforms and one substrate-inducible nitrate reductase (iNR) isoform in leaves. All three NR isoforms are expressed in cv. Williams 82 while the nr1 mutant expresses only the iNR isoform. The genetic and molecular mechanisms for regulation of these isoforms have not been elucidated. We describe here the isolation, by reverse transcription-polymerase chain reaction (RT-PCR), of two cDNA clones encoding soybean NR. They were designated as iNR1 and iNR2, respectively, since both were inducible by nitrate. The iNR1 and iNR2 cDNAs cover total encoding regions of 2661 and 2673 nucleotides, respectively. The iNR1 clone shows a 12 bp deletion at the 5' end, relative to iNR2. They show overall similarity of 89% at the nucleotide level, and 87% at the amino acid level. Like all plant NRs cloned so far, deduced amino acid sequences between iNR1 and iNR2 show greatest variation at the N-terminal region while no difference was observed at the C-terminus. Soybean iNR mRNAs were found to be different from those of maize and tobacco in response to tungsten inhibitor treatment, since the inhibitor decreased the steady-state levels of mRNA for soybean iNR and for NiR. Using the same 5' regions of both cDNAs as the probes, Southern blot analysis of genomic DNA revealed differences in organization between iNR1 and iNR2. The genomic DNA from wild-type Williams 82 soybean was shown to have three Eco RI fragments while the nr1 mutant lacked an 8 kb fragment when probed with iNR1 cDNA. Likewise, the nr1 mutant lacked three Hae III restriction fragments when probed with iNR1 cDNA. When probed with iNR2, both wildtype and nr1 mutant showed one identical Eco RI band and two identical Hae III bands. In northern blots, the steady-state level of iNR1 mRNA was similar for the nr1 mutant and the wild-type parent after 20 to 48 h induction by nitrate. Based on the Eco RI and Hae III restriction enzyme digestion patterns observed in Southern blot analysis of soybean DNA, it is concluded that in soybean iNR1 is encoded by a small multiple gene family and iNR2 is a single gene.
  Plant Molecular Biology 12/1995; 29(3):491-506. · 4.15 Impact Factor
• Source

  Available from: Randy Scholl

  Article: Improvements of the nitrite color development in assays of nitrate reductase by phenazine methosulfate and zinc acetate.

  R L Scholl, J E Harper, R H Hageman
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  ABSTRACT: Nitrate reductase activity is most commonly assayed by measurement of product formation. Excess NADH and factor(s) present in the enzyme extract that interfere with the diazotization and azo color complex of nitrite cause a depression of apparent nitrate reductase activity. Two postassay treatments were found that markedly enhanced the extent of nitrite color formation and apparent nitrate reductase activity. The procedure involves stopping the reaction with zinc acetate (50 mumoles per ml of reaction mix), followed by removal of the precipitate by centrifugation. Presumably the zinc acetate removes extract factor(s) that interfere with color development, because it does not remove the NADH. Phenazine methosulfate (15 nmoles per ml of reaction mix) is added to aliquots of the supernatant and allowed to stand for 20 min at 30 C to oxidize the residual NADH before color development.
  Plant physiology 07/1974; 53(6):825-8. · 6.53 Impact Factor
• Source

  Available from: plantphysiol.org

  Article: Canopy and Seasonal Profiles of Nitrate Reductase in Soybeans (Glycine max L. Merr.).

  J E Harper
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  ABSTRACT: Nitrate reductase activity of soybeans (Glycine max L. Merr.) was evaluated in soil plots and outdoor hydroponic gravel culture systems throughout the growing season. Nitrate reductase profiles within the plant canopy were also established. Mean activity per gram fresh weight per hour of the entire plant canopy was highest in the seedling stage while total activity (activity per gram fresh weight per hour times the total leaf weight) reached a maximum when plants were in the full bloom to midpod fill stage. Nitrate reductase activity per gram fresh weight per hour was highest in the uppermost leaf just prior to full expansion and declined with leaf position lower in the canopy. Total nitrate reductase activity per leaf was also highest in the upper-most fully expanded leaf during early growth stages. Maximum total activity shifted to leaf positions lower in the plant canopy with later growth stages.Nitrate reductase activity of soybeans grown in hydroponic systems was significantly higher than activity of adjacent soil grown plants at later growth stages, which suggested that under normal field conditions the potential for nitrate utilization may not be realized. Nitrate reductase activity per gram fresh weight per hour and nitrate content were positively correlated over the growing season with plants grown in either soil or solution culture. Computations based upon the nitrate reductase assay of plants grown in hydroponics indicated that from 1.7 to 1.8 grams N could have been supplied to the plant via the nitrate reductase process. The harvested seed contained 1.1 to 1.2 grams N per plant. Thus, based on previous estimates of approximately 32% of the final N distribution being in the vegetative plant parts, the estimated input of reduced nitrogen via the enzyme assay was in agreement with the actual N accumulation.The amount of calculated N(2)-fixation by nodules per season with plants grown in hydroponics was less than 2% of the computed nitrate reduced via leaf nitrate reductase. Thus, the level of nitrate in the nutrient solution appeared to be quite inhibitory to N(2)-fixation.
  Plant physiology 03/1972; 49(2):146-54. · 6.53 Impact Factor
• Article: Improvements oftheNitrite ColorDevelopment inAssays ofNitrate Reductase byPhenazine Methosulfate

  R. L. SCHOLL, J. E. HARPER, H. HAGEMA
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  ABSTRACT: Nitrate reductase activity ismostcommonly assayed bymea- surementofproduct formation. Excess NADH andfactor(s) present intheenzymeextract thatinterfere withthediazotiza- tionandazocolorcomplex ofnitrite causea depression of apparent nitrate reductase activity. Two postassay treatments werefoundthatmarkedly enhanced theextentofnitrite color formation andapparent nitrate reductase activity. Theproce- dureinvolves stopping thereaction withzincacetate(50 ,umoles perml ofreaction mix),followed byremoval ofthe precipitate bycentrifugation. Presumably thezincacetatere- movesextract factor(s) thatinterfere withcolor development, because itdoesnotremovetheNADH.Phenazine methosulfate (15nmoles permlofreaction mix)isaddedtoaliquots ofthe supernatant andallowed tostandfor20minat30C tooxidize theresidual NADH before color development. andUnited States Regional Soybean Laboratory, Urbana,