We compared two putative Freeman maples [`Jeffersred', (Autumn Blaze ®) and `Indian Summer'] and five red maples [`Franksred' (Red Sunset ®), `Autumn Flame', `PNI 0268' (October Glory®), `Fairview Flame', and unnamed selection 59904] for effects of flooding on stomatal conductance. A method for quantifying changes in leaf color that occurred on flooded plants also was developed. Potted plants grown from rooted cuttings in a greenhouse were subjected to 75 days of root-zone inundation (flood treatment) or were irrigated frequently (control treatment). Across genotypes, stomatal conductance of flooded plants initially increased by about 20% and then fell to and was sustained below 50 mmol·s–1·m–2. Stomatal conductance of flooded plants of `Indian Summer' decreased to 20 mmo·s–1·m–2 after 8 days of inundation, and two of three flooded `Indian Summer' plants died during treatment. Other genotypes required at least twice this time to display a similar reduction in stomatal conductance, indicating `Indian Summer' may be particularly flood sensitive. Intensities of red, green, and blue color at a consistent interveinal position were analyzed with Visilog software by using scanned leaf images of the youngest fully expanded leaf of each plant in both treatments. A genotype × irrigation interaction existed for the ratio of green to red intensity. This method provided numerical data that corresponded well to differences among genotypes we observed visually. For example, while flooding did not alter the color of `Autumn Flame' leaves, the ratio of green to red was three times greater for controls of Autumn Blaze® than for the flooded plants of this cultivar.
James A. Zwack, William R. Graves, and Alden M. Townsend
J. Ryan Stewart, William R. Graves, and Reid D. Landes
Can Carolina buckthorn (Rhamnuscaroliniana) persist north of its native habitat without becoming invasive? Its distribution (USDA zones 5b to 9b) suggests that genotypes vary in cold hardiness, and invasiveness of other Rhamnus sp. has been linked to unusually early budbreak each spring. Therefore, we investigated depth of cold hardiness and vernal budbreak of Carolina buckthorns from multiple provenances and made comparisons to the invasive common buckthorn (Rhamnus cathartica). Budbreak was recorded in Ames, Iowa, from 9 Apr. to 10 May 2002. Buds of common buckthorn broke earlier than those of Carolina buckthorn, and mulching plants of Carolina buckthorn hastened budbreak. Stem samples were collected in October, January, and April from a plot in Ames, Iowa (USDA zone 5a), of Carolina buckthorns from three provenances (Missouri, Ohio, and Texas) and of naturalized common buckthorns. A similar schedule was followed during the next winter, when two plot locations [Ames, Iowa, and New Franklin, Mo. (USDA zone 5b)], were compared, but Carolina buckthorns from only Missouri and Texas were sampled. Carolina buckthorn and common buckthorn survived midwinter temperatures as low as –21 °C and –24 °C, respectively. Provenance differences were minimal; Carolina buckthorns from Missouri were more hardy than those from Ohio and Texas only in April of the first winter. We conclude that its cold hardiness will permit use of Carolina buckthorn beyond where it is distributed in the southeastern United States. Delayed budbreak of Carolina buckthorn relative to that of common buckthorn may underscore the potential for Carolina buckthorn in regions with harsh winters and may lessen its potential to be as invasive as common buckthorn.
Michael S. Dosmann, William R. Graves, and Jeffery K. Iles
The limited use of the katsura tree (Cercidiphyllum japonicum Sieb. & Zucc.) in the landscape may be due to its reputed, but uncharacterized, intolerance of drought. We examined the responses of katsura trees subjected to episodes of drought. Container-grown trees in a greenhouse were subjected to one of three irrigation treatments, each composed of four irrigation phases. Control plants were maintained under well-hydrated conditions in each phase. Plants in the multiple-drought treatment were subjected to two drought phases, each followed by a hydration phase. Plants in the single-drought treatment were exposed to an initial drought phase followed by three hydration phases. Trees avoided drought stress by drought-induced leaf abscission. Plants in the multiple- and single-drought treatments underwent a 63% and 34% reduction in leaf dry weight and a 60% and 31% reduction in leaf surface area, respectively. After leaf abscission, trees in the single-drought treatment recovered 112% of the lost leaf dry weight within 24 days. Leaf abscission and subsequent refoliation resulted in a temporary reduction in the leaf surface area: root dry weight ratio. After relief from drought, net assimilation rate and relative growth rate were maintained at least at the rates associated with plants in the control treatment. We conclude that katsura is a drought avoider that abscises leaves to reduce transpirational water loss. Although plants are capable of refoliation after water becomes available, to maintain the greatest ornamental value in the landscape, siting of katsura should be limited to areas not prone to drought.
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).
Hongyi Zhang, William R. Graves, and Alden M. Townsend
We determined transpiration rate, survival, and rooting of unmisted, softwood cuttings of `Autumn Flame' red maple (Acer rubrum L.) and `Indian Summer' Freeman maple (Acer ×freemanii E. Murray). Effects of perlite at 24, 30, and 33 °C were assessed to determine whether responses of cuttings would be consistent with cultivar differences in resistance to root-zone heat previously shown with whole plants. During 7 d, cutting fresh mass increased by ≈20% at all temperatures for `Autumn Flame' red maple, but fresh mass of `Indian Summer' Freeman maple decreased by 17% and 21% at 30 and 33 °C, respectively. The percentage of cuttings of `Indian Summer' that were alive decreased over time and with increasing temperature. Transpiration rate decreased during the first half of the treatment period and then increased to ≈1.1 and 0.3 mmol·m-2·s-1 for `Autumn Flame' and `Indian Summer', respectively. Mean rooting percentages over temperatures for `Autumn Flame' and `Indian Summer' were 69 % and 16%, respectively. Mean rooting percentages at 24, 30, and 33 °C over both cultivars were 74%, 29%, and 25%, respectively. Over temperatures, mean root count per cutting was 41 and seven, and mean root dry mass per cutting was 4.9 and 0.4 mg, for `Autumn Flame' and `Indian Summer', respectively. Use of subirrigation without mist to root stem cuttings was more successful for `Autumn Flame' than for `Indian Summer'. Temperature × cultivar interactions for cutting fresh mass and the percentage of cuttings remaining alive during treatment were consistent with previous evidence that whole plants of `Autumn Flame' are more heat resistant than plants of `Indian Summer'. Mass and survival of stem cuttings during propagation in heated rooting medium may serve as tools for screening for whole-plant heat resistance among maple genotypes.
Olivia M. Lenahan, William R. Graves, and Rajeev Arora
Styrax americanus Lam. (American snowbell) is a deciduous shrub or small tree seldom produced in nurseries. This species is distributed in patchy populations found mainly from Florida to southern Illinois, although a small, disjunct population exists in northern Illinois. The winter-hardiness and loss of hardiness during a period of increased temperature (deacclimation) of plants from this disjunct population may differ from those of S. americanus elsewhere. We examined cold-hardiness and deacclimation of stems of plants from the disjunct population, from southern Illinois, and from Florida. Segments of stems removed from plants grown outdoors in Ames, IA, were exposed to low-temperature ramping, and the temperature at which stems showed 50% damage (LT50) was determined by using the tissue-discoloration method. To assess deacclimation, stem segments were collected from cold-acclimated plants during winter in a minimally heated greenhouse and exposed to controlled warm temperatures for various time intervals followed by low-temperature ramping. Plants from Illinois were ≈15 °C more cold-hardy than plants from Florida in Feb. 2008. Plants from the disjunct population in northern Illinois showed less stem tip injury than did plants from southern Illinois. Deacclimation patterns were similar between plants from both Illinois populations. Plants sampled in Apr. 2009 from Florida deacclimated more rapidly than corresponding samples from Illinois, and the chilling required to overcome endodormancy increased with increasing latitude of plant origin. This research suggests that germplasm from the Illinois populations should be used in regions where the poorer hardiness and deacclimation resistance of most S. americanus would not permit survival.
Kimberly A. Klock, Henry G. Taber, and William R. Graves
Horticultural species vary in growth response to high root-zone temperature (RZT), but little is known about the effects of RZT on nutrient uptake. We determined P, K, Ca, Mg, Zn, and Mn total plant content of tomato (Lycopersicon esculentum Mill. cv. Jet Star), muskmelon (Cucumis melo L. cv. Gold Star), and honey locust (Gleditsia triacanthos L. var. inermis Willd.) grown in nutrient solution kept at 24, 27, 30, 33, and 36C. RZT effects on plant dry mass gain and gain in nutrient per plant varied by species. Honey locust and tomato total plant gain in P decreased linearly with increasing RZT, while melon P content increased linearly to 36C. Trends in total Mg and Mn content will be presented, as well as results from further research on correlations between supraoptimal RZT, root respiration, and shoot and root P content of tomato.
Carol M. Foster, William R. Graves, and Harry T. Horner
Knowing whether leguminous trees have the potential to nodulate after infection by rhizobial bacteria is important for managing nitrogen (N) applications during tree production and for culture in the landscape. Although 98% of studied species in the Papilionoideae nodulate, the nodulation status of two tree species in this subfamily is uncertain. Cladrastis kentukea (Dum.-Cours.) Rudd (American yellowwood) did not form nodules during inoculation studies in 1939 and 1992. Nodules were observed on mature Sophora japonica L. (Japanese pagodatree) in Japan and Hawaii in the 1940s, but compatible rhizobia reportedly isolated in Japan are no longer held in bacterial collections. Our objective was to verify further that American yellowwood does not nodulate and to confirm reports that Japanese pagodatree does nodulate. Rhizobia that infect many plant hosts, soil samples and rhizobial isolates from other Sophora spp., and soil samples from mature American yellowwood and Japanese pagodatree were used to inoculate 5-day-old seedlings of American yellowwood, Japanese pagodatree, and control species. Soil from indigenous and introduced trees in the continental United States, Hawaii, Japan, and China was used. Inoculated and uninoculated plants were grown for 7 weeks in sterile Leonard jars or clay pots containing perlite and irrigated with sterile, N-free Hoagland's solution. No inoculation treatment elicited nodulation of American yellowwood or Japanese pagodatree. Our results provide additional evidence that American yellowwood lacks that capacity to nodulate and cast further doubt on nodulation of Japanese pagodatree.
Janet McCray Batzli, William R. Graves, and Peter van Berkum
Our objectives were to test whether Maackia amurensis Rupr. & Maxim. nodulates and fixes N and to characterize the N-fixing bacteria effective with this host. Soil samples were collected near diverse legume trees at arboreta and public gardens in the United States, Canada, and China. Seedlings of M. amurensis were grown for 6 weeks in a low-N, sterile medium and inoculated with soil samples. At harvest, nodules were found on the lateral and upper portions of root systems. Bacteria were isolated from nodules and subculture. Roots of seedlings inoculated with all 11 of these isolates nodulated and freed N, confirming that the isolates were rhizobial bacteria. Growth of isolates in axenic culture generally was poor when single sources of C were provided. Generation times of the isolates ranged from 6 to 10 hours, and all isolates raised the pH of culture media. Isolates were highly resistant to several antibiotics, showed no 6-phosphogluconate dehydrogenase (6PGD) or β-galactosidase activity, and were highly sensitive to NaCl. These results provide the first evidence that M. amurensis has the capacity to form N-fixing symbioses with rhizobial bacteria and indicate that the bacteria are Bradyrhizobium sp.
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.