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Carol M. Foster, William R. Graves, and Harry T. Horner

ENOD2 and other early nodulin genes are conserved among legumes studied to date and might function as markers for the potential of legumes to nodulate. Early nodulin genes have been characterized only among herbaceous legumes. We are interested in understanding the nature of ENOD2 in a nodulating, woody legume. A 561-bp MaENOD2 PCR fragment was used as a probe to screen a cDNA library from nodules ≈1 mm in diameter on roots of Amur maackia, the only temperate and horticulturally desirable leguminous tree species known to nodulate. Five cDNAs were selected for nucleotide sequence analysis. Sequences were determined by using automated dideoxy sequencing and analyzed for identity to other genes with the Genetics Computer Group (GCG) program. The cDNA clones show 68% to 74% identity at the nucleic acid level with ENOD2 genes of Sesbania rostrata Brem. & Oberm., Glycine max (L.) Merrill, and Lupinus luteus L. Southern and northern analyses are being conducted to investigate the possibility of a gene family and to show differential and temporal production of transcripts, respectively. These studies provide new information about nodulins of woody legumes and are being used to facilitate related research on molecular barriers to nodulation in the closely related, non-nodulating tree species Cladrastis kentukea (Dum.-Cours.) Rudd (American yellowwood) and Sophora japonica L. (Japanese pagodatree).

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Carol M. Foster, William R. Graves, and Harry T. Horner

A complete picture of legume nodulation has yet to be elucidated. Most studies of the molecular mechanisms responsible for nodule organogenesis have focused on herbaceous legumes. We investigated the presence of a putative ENOD2 gene and studied the temporal and organ-specific production of its transcripts in an ornamental woody legume, Amur maackia. Primers derived from proline-rich pentapeptide repeats of conserved ENOD2 sequences and the genomic DNA of Amur maackia were used to obtain a 543-bp PCR fragment. Southern and Northern blots were probed with this cloned fragment. The Amur maackia genome contained an ENOD2 sequence that is similar to sequences in other species. Expression of the putative ENOD2 gene was detected in roots, 4 days after rhizobial inoculation, but not in leaves or stems. New data on the characteristics of nodulin genes in woody legumes will be beneficial in clarifying the nature and evolution of nodulation in legumes and may have implications for developing sustainable nursery production protocols.

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Carol M. Foster, William R. Graves, and Harry T. Horner

Early nodulin genes, such as ENOD2, may be conserved and could function as molecular markers for nodulation. Many nodulating and nonnodulating legumes must be analyzed before the role of such genes in nodulation can be determined. Japanese pagodatree and American yellowwood are closely related, ornamental woody legumes. Unsubstantiated reports of nodulation in Japanese pagodatree require confirmation, and American yellowwood has not been observed to nodulate. We investigated the presence of putative ENOD2 genes in these species, and we are studying differential and temporal expression. Genomic DNA of Japanese pagodatree and primers, derived from proline-rich pentapeptide repeats of conserved ENOD2 sequences, were used to obtain a 555-bp PCR fragment. This cloned fragment was used as a probe for Southern and Northern hybridizations. Genomes of Japanese pagodatree and American yellowwood contained sequences that are similar to ENOD2 sequences in other legumes. Treatments with either cytokinin or an auxin transport inhibitor may induce expression of the putative ENOD2 genes. New data on the characteristics of nodulin genes in woody legumes will clarify the nature and evolution of nodulation in legumes and may have implications for developing sustainable nursery production protocols.

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Yuejin Weng, Jun Qin, Stephen Eaton, Yufeng Yang, Waltram Second Ravelombola, and Ainong Shi

Cowpea [ Vigna unguiculata (L.) Walp], an annual legume native to tropical and subtropical regions, is a protein-rich crop that complements staple cereal for human and fodder for livestock and also provides soil improvement benefits through

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Patrick Chesney, Linda Wessel-Beaver, and Donald N. Maynard

Most cultivars of tropical pumpkin (Cucurbita moschata Duchesne) are large, trailing plants. New semi-bush (SB) genotypes need to be tested against traditional long vine (LV) types. Both types of pumpkin have large amounts of interplant space during the early stages of growth that might allow for the planting of an intercrop. To test this hypothesis, as well as the performance of tropical pumpkins of varying growth habit, double rows of beans (Phaseolus vulgaris L.) or cowpeas [Vigna unguiculata (L.) Walp.] were intercropped between rows of SB or traditional LV tropical pumpkin in Spring and Fall 1993 in Lajas and Isabela, Puerto Rico. In general, interactions between intercrop treatment and pumpkin genotype were not significant. At its maximum percentage cover (MC) the LV genotype covered, or nearly covered, the entire plot while the SB genotype covered 50% of the plot or less. The SB pumpkin was harvested 5 to 27 days earlier than the LV type. Yield was two to 12 times greater, and average fruit size three to six times greater in the latter. Planting of an intercrop did not reduce pumpkin yield. Green-shelled yields of intercropped legumes averaged ≈700 kg·ha-1. Genotype of the pumpkin maincrop did not affect legume green-shelled yields in Lajas. In Isabela, legume green-shelled yields were 50% higher in SB than in LV pumpkin plots. Legume dry grain yields were greatly reduced in LV compared to SB plots. Intercropping of tropical pumpkin with a short season legume that can be harvested green-shelled is an efficient intercropping system that offers additional yield from the legume without sacrificing yield from the pumpkin maincrop. Both SB and LV pumpkins can be used in an intercrop system, but pumpkin yields were much greater with the LV genotype.

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Rodney Jones and Robert Geneve

Redbud (Cercis canadensis) is a small woody ornamental legume that has a hard seed coat, which imposes physical dormancy, typical of many legumes. Redbud also possesses an internal embryo dormancy that must be overcome by stratification. In order to observe the relationship between anatomy and germination, seeds were embedded in JB-4 resin during various developmental and germination stages. The seeds were cut longitudinally with a glass bladed microtome, to observe the radicle, vascular traces and testa. It appears that the vascular traces left from the funiculus serve as a weak point in non-dormant seeds that allows the radicle to rupture the testa during germination.

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William R. Graves and Lorna C. Wilkins

A laboratory exercise for illustrating aspects of biological nitrogen fixation (BNP) to students in plant science courses is described. Surface-sterilized seeds of black locust (Robinia pseudoacacia L.) and soybean (Glycine max Merill) were sown together in plastic containers filled with a sterile, soilless medium. Containers were assigned randomly to treatments designed to show how inoculation with two strains of rhizobial bacteria and application of nitrate affect root nodulation and plant growth. Results demonstrated that BNF occurs in diverse legumes, that legumes vary in the strains of rhizobia with which they associate, that nodulation is inhibited by nitrate, and that dependency on BNP can reduce growth compared with plants provided nitrate.

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Bharat P. Singh, Upendra M. Sanju, and Wayne F. Whitehead

Our objective was to determine the effect of winter cover crops on the yield and N concentration of the following crop of tomato. No commercial fertilizer was applied to the tomato crop. Cover crops were planted in fall in a randomized complete-block design with control (fallow), rye, hairy vetch, and crimson clover treatments. `Mountain Pride' tomato was planted in spring after incorporating cover crops into the soil. Soil inorganic N content during the tomato growing season was significantly affected by the nature of cover crops planted during winter. Tomato planted after legumes had significantly greater amounts of inorganic N available for uptake compared to nonlegume or control. A rye cover crop did not have any effect on the yield of the ensuing tomato crop. On the contrary, a 15% increase in tomato fruit yields resulted from cover cropping with legumes. The N concentration in fruit in all treatments was similar. However, tomato grown after rye had significantly lower vegetative N concentration. Total N uptake was significantly greater in tomato succeeding legumes compared to nonlegume or fallow. It was concluded that by adding inorganic N into the soil, legumes increased the fruit yield and N uptake of the succeeding tomato crop.

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Wilfred Singogo, William J. Lamont Jr., and Charles W. Marr

Four cover crops {alfalfa (Medicago sativa L. `Kansas Common'), hairy vetch (Vicia villosa Roth), Austrian winter pea [Pisum sativum subsp. arvense (L.) Poir], and winter wheat (Triticum aestivum L. `Tam 107')}, alone and in combination with feedlot beef manure at 5 t·ha–1 were evaluated for 2 years to determine whether sufficient N could be supplied solely by winter cover cropping and manure application to produce high-quality muskmelons (Cucumis melo L. `Magnum 45') in an intensive production system using plastic mulch and drip irrigation. Among the legumes, hairy vetch produced the most biomass (8.9 t·ha–1) and accumulated the most N (247 kg·ha–1). Winter wheat produced more biomass (9.8 t·ha–1) than any of the legumes but accumulated the least N (87 kg·ha–1). Melon yields produced using legume cover crops alone were similar to those receiving synthetic N fertilizer at 70 or 100 kg·ha–1. Melons produced on plots with cover crops combined with beef manure did not differ significantly in yield from those produced on plots with only cover crops. Legume cover crops alone, used with plastic mulch and drip irrigation, provided sufficient N for the production of high-quality muskmelons.

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John R. Teasdale and Aref A. Abdul-Baki

Hairy vetch (Vicia villosa Roth), crimson clover (Trifolium incarnatum L.), and rye (Secale cereale L.) and mixtures of rye with hairy vetch and/or crimson clover were compared for no-tillage production of staked, fresh-market tomatoes (Lycopersicon esculentum Mill.) on raised beds. All cover crops were evaluated both with or without a postemergence application of metribuzin for weed control. Biomass of cover crop mixtures were higher than that of the hairy vetch monocrop. Cover crop nitrogen content varied little among legume monocrops and all mixtures but was lower in the rye monocrop. The C:N ratio of legume monocrops and all mixtures was <30 but that of the rye monocrop was >50, suggesting that nitrogen immobilization probably occurred only in the rye monocrop. Marketable fruit yield was similar in the legume monocrops and all mixtures but was lower in the rye monocrop when weeds were controlled by metribuzin. When no herbicide was applied, cover crop mixtures reduced weed emergence and biomass compared to the legume monocrops. Despite weed suppression by cover crop mixtures, tomatoes grown in the mixtures without herbicide yielded lower than the corresponding treatments with herbicide in 2 of 3 years. Chemical name used: [4-amino-6-(1,1-dimethylethyl)-3-(methylthio)-1,2,4-triazin-5(4H)-one](metribuzin).