Nodulation of Seaside Alder Topdressed with Controlled-release Fertilizer

Seaside alder (Alnus maritima) is a rare species adapted for ornamental use. It can be produced easily by commercial growers, and when soil-inoculated in nursery containers, will form root nodules containing nitrogen (N)-fixing bacteria in the genus Frankia. It has been demonstrated that nodulation within this species can be controlled by the amount of N plants receive from a liquid source; however, granular controlled-release fertilizers (CRF) also are commonly used by commercial growers, and information is not available on CRF effects on nodulation. We sought to determine the application rate of CRF that will maximize nodulation of seaside alder while sustaining acceptable plant health and growth. Inoculated containerized plants were topdressed with CRF at eight application rates from 0 to 32.0 g per container. A control group of uninoculated plants received the manufacturer's prescribed rate of 6.0 g per 6-inch-diameter standard round pot. Regression analysis revealed that treatment with 2.0 g of CRF maximized root nodule formation (a mean of 26 nodules), while nodulation was inhibited at CRF rates ≥4.0 g. Plant growth parameters and leaf N content of inoculated plants treated with 2.0 g of CRF were similar to those of uninoculated plants receiving the prescribed rate of CRF (P < 0.05); nitrate-nitrogen leaching from pots containing 2.0-g CRF-treated plants was minimal. We conclude that vigorous nodulated plants of seaside alder can be produced, with minimal nitrate leaching, by providing CRF at lower than prescribed rates.

Systematics of Alnus maritima (Seaside Alder) Resolved by ISSR Polymorphisms and Morphological Characters

Alnus maritima (Marsh.) Muhl. ex Nutt. is a rare woody plant species that exists as three subspecies found in widely disjunct locations in the United States. Although there is a growing interest in the phytogeography, ecology, conservation, and landscape potential of this species, the phylogeny of A. maritima has not yet been resolved by using molecular methods. We have combined a relatively new method of genome fingerprinting, ISSR-PCR, and the automated imaging capabilities of GeneScan technology to investigate the molecular systematics of A. maritima. Based on the molecular evidence from 108 ISSR loci, we confirm that the three disjunct populations of A. maritima have diverged sufficiently to be classified as subspecies. Our molecular phylogeny of the three subspecies of A. maritima agreed in topology with a phylogeny produced from morphological data and showed that subsp. oklahomensis is the most distinct of the three subspecies and was the first to diverge. The simultaneous analysis of molecular and morphological data provides a detailed and balanced phylogeny reconstruction for the three subspecies. Our results support the theory that A. maritima originated in Asia, migrated into North America across the Bering land bridge, and was established over a large range in the New World before being forced into its present meager disjunct distribution.

041 Dehydrins as Drought-stress Mechanisms in Leaves of Red Maple and Seaside Alder

Desiccation damage in ornamental plant species is of particular concern to the nursery and landscape industry. Species in two genera, Acer and Alnus, display fundamental differences in how drought affects leaves. The same soil moisture content that causes foliar desiccation and abscission in Alnus maritima (Marsh.) Nutt. (seaside alder) causes neither response in Acer rubrum L. (red maple). Understanding molecular mechanisms associated with plant response to drought stress can be an im portant factor in developing strategies for improved sustainability in urban landscapes. Our objective was to characterize expression of drought-induced dehydrin genes in leaves of `Red Sunset' red maple (desiccation-resistant) and seaside alder (desiccation-sensitive) in response to dehydration and rehydration. Potted cuttings grown in a glasshouse were subjected to four cycles of drought and rehydration. Stomatal conductance and volumetric moisture content of rooting medium were used to determine when drought cycles ended. During the second and fourth cycles, leaves were sampled for RNA and protein extraction. Dehydrin probes were generated from genomic DNA of both species by using PCR with primers designed from conserved regions in dehydrin genes. Southern blot analyses revealed the presence of dehydrin genes in seaside alder and red maple genomes. Reverse transcriptase (RT)-PCR was used to isolate desiccation-induced dehydrin cDNAs from total RNA extracted from drought-stressed leaves. The cDNA clones show 61% to 66% identity at the nucleic acid level with dehydrin genes of soybean, sunflower, radish, and potato. Accumulation of dehydrin transcripts and proteins in leaves in response to dehydration and rehydration are being studied through northern and western blot analyses, respectively. Our results may lead to a rapid screening technique for seedlings with improved mechanisms of drought resistance.

143 Gas Exchange, Water Relations, and Growth of Seaside Alder Exposed to Drought and Flooding

Alnus maritima [Marsh.] Nutt. (seaside alder) is a rare species that occurs naturally only on soils that are frequently or constantly saturated with fresh water. The objective of our first experiment was to determine effects of drought and flooding treatments of differing severity on foliar gas exchange, water relations, and development of plants grown in containers in a greenhouse. In a second experiment we examined how the rate of water loss from soil during drought episodes affected the gas exchange and survival of leaves. In the first experiment, changes in soil moisture content, which ranged from saturation to 10% or less by volume across treatments, were associated with altered stem water potential and net photosynthesis. Analysis of the osmolarity of liquid extracted from leaves indicated that osmotic adjustment did not occur in response to drought. Shoot dry weight per plant ranged from over 7 g (only the lower portion of the soil profile kept saturated) to less than 3 g (entire soil profile constantly saturated). Episodes of drought of different severity led to plants with shoots that weighed between these two extremes, and exposure to soils with 10% water or less by volume did not elicit leaf desiccation or abscission. Results of the second experiment suggest that leaf desiccation can result from exposing plants to 10% water or less by volume if the drought develops rapidly in a small volume of soil. We conclude that, despite the niche it occupies in nature, seaside alder may have the potential to be used in managed landscapes with soils that vary in moisture content.

Propagation of Alnus maritima from Softwood Cuttings

Alnus maritima (Marsh.) Nutt. (seaside alder) is a rare, woody-plant species with potential for use in managed landscapes. Information on the propagation and production of this species is not available. Our objective was to evaluate the use of softwood cuttings to propagate A. maritima, with emphasis on how indole-3-butyric acid (IBA), plant provenance, and time of collection affect cutting survival, rooting percentage, the number of roots produced, and their length. Propagation trials were conducted with cuttings from seven trees on the Delmarva Peninsula (Eastern Shore of Maryland and southern Delaware) and seven trees in Oklahoma. Cuttings from mature plants in both provenances were collected on 14 June and 23 Aug. 1998; wounded; treated with IBA at 0, 1, or 8 g·kg-1; and placed under intermittent mist in a greenhouse for 9 weeks. Use of IBA at 8 g·kg-1 caused a greater rooting percentage (68%), root count (7.2), and root length (39.2 mm) than did the other IBA rates when applied to cuttings from Oklahoma in June, but IBA had little effect on cuttings from the Delmarva Peninsula. Across IBA treatments, rooting of cuttings from Oklahoma (55% in June and 12% in August) was greater than the rooting of cuttings from Delmarva (27% in June and 3% in August). Cuttings from Oklahoma had greater survival, callus development, root length, and root count than did cuttings from the Delmarva Peninsula during June and August trials. Averaged over IBA treatments and provenances, cuttings collected on 14 June rooted more frequently (41%) than did cuttings collected 23 Aug. (8%). We conclude that softwood cuttings from mature plants are an effective way to multiply clones of A. maritima, particularly when cuttings are collected early in the season and treated with IBA at 8 g·kg-1.