Richard L. Harkess

Poplar maintains zinc homeostasis with heavy metal genes HMA4 and PCS1

Perennial woody species, such as poplar (Populus spp.) must acquire necessary heavy metals like zinc (Zn) while avoiding potential toxicity. Poplar contains genes with sequence homology to genes HMA4 and PCS1 from other species which are involved in heavy metal regulation. While basic genomic conservation exists, poplar does not have a hyperaccumulating phenotype. Poplar has a common indicator phenotype in which heavy metal accumulation is proportional to environmental concentrations but excesses are prevented. Phenotype is partly affected by regulation of HMA4 and PCS1 transcriptional abundance. Wild-type poplar down-regulates several transcripts in its Zn-interacting pathway at high Zn levels. Also, overexpressed PtHMA4 and PtPCS1 genes result in varying Zn phenotypes in poplar; specifically, there is a doubling of Zn accumulation in leaf tissues in an overexpressed PtPCS1 line. The genomic complement and regulation of poplar highlighted in this study supports a role of HMA4 and PCS1 in Zn regulation dictating its phenotype. These genes can be altered in poplar to change its interaction with Zn. However, other poplar genes in the surrounding pathway may maintain the phenotype by inhibiting drastic changes in heavy metal accumulation with a single gene transformation.

Shoot morphogenesis associated with flowering in Populus deltoides (Salicaceae)

Temporal and spatial formation and differentiation of axillary buds in developing shoots of mature eastern cottonwood (Populus deltoides) were investigated. Shoots sequentially initiate early vegetative, floral, and late vegetative buds. Associated with these buds is the formation of three distinct leaf types. In May of the first growing season, the first type begins forming in terminal buds and overwinters as relatively developed foliar structures. These leaves bear early vegetative buds in their axils. The second type forms late in the first growing season in terminal buds. These leaves form floral buds in their axils the second growing season. The floral bud meristems initiate scale leaves in April and begin forming floral meristems in the axils of the bracts in May. The floral meristems subsequently form floral organs by the end of the second growing season. The floral buds overwinter with floral organs, and anthesis occurs in the third growing season. The third type of leaf forms and develops entirely outside the terminal buds in the second growing season. These leaves bear the late vegetative buds in their axils. On the basis of these and other supporting data, we hypothesize a 3-yr flowering cycle as opposed to the traditional 2-yr cycle in eastern cottonwood.

Structure and developmental regulation of CONSTANS-LIKE genes isolated from Populus deltoides

Abstract Two CONSTANS - LIKE ( COL ) genes, PdCO1 and PdCO2 , were isolated from eastern cottonwood ( Populus deltoides Bartr. ex. Marsh. var. deltoides ). PdCO1 is unique among all the characterized CONSTANS-LIKE genes in the plant kingdom in that the deduced protein sequence contains a putative signal peptide with a potential transmembrane helix at the N-terminal end. Current bioinformatics tools suggest that PdCO1 protein may be secreted. Immuno-blot analysis indicated that although the PdCO1 was abundant in leaf, it was not detected in the subtended bud. Temporal mRNA and protein expression of PdCO1 and PdCO2 was determined in the leaf and bud at the ninth node of shoots, where transition from vegetative to floral phase has been observed to occur. The expression of both genes in the leaf increased dramatically from March 30 to April 21 and decreased on May 9. The increased expression coincided with morphological changes occurring in the axillary meristem of the subtended bud and with increased abundance of PdCO1 protein in the leaf. These changes sequentially included (1) the cessation of leaf-primordium formation, (2) the transition of the axillary shoot apical meristem to an inflorescence meristem, (3) and the subsequent formation of bracts and floral meristems in the axils of bracts flanking the inflorescence meristem. The temporal expression of PdCO1 and PdCO2 prior to flower bud development suggests that these genes might be involved in floral induction.

Protein source and nutrient density in the diets of male broilers from 8 to 21 d of age: Effects on small intestine morphology

In a companion study, high amino acid (AA) or apparent metabolizable energy (AME) densities in the diets of broilers from 8 to 21 d of age were found to improve feed conversion. A total of 1,120 male Ross×Ross 708 chicks were randomly allocated to 80 pens (8 treatments, 10 replications per treatment, 14 chicks per pen). A 2×2×2 factorial arrangement of treatments was used to investigate the interaction among the protein source (high distillers dried grains with solubles diet [hDDGS] or high meat and bone meal diet [hMBM]), AA density (moderate or high), and AME density (2,998 or 3,100 kcal/kg) of diets on small intestine morphology. Duodenum, jejunum, and ileum samples from 2 chicks per pen were collected and measured individually at 21 d. Jejunum sections were processed for histological analysis. Chicks fed hDDGS diets exhibited longer small intestines than did chicks fed hMBM diets. Particularly, when chicks were fed high AA density diets, jejuna were longer in groups fed hDDGS diets than groups fed hMBM diets. Dietary treatments did not affect jejunum villus height, width, area, crypt depth, villus to crypt ratio, goblet cell size, or cell density. In birds fed diets containing a moderate AA and a high AME density, jejunum muscle layers of chicks fed hDDGS diets were thicker than those fed hMBM diets. Chicks exhibited a lower feed conversion ratio (FCR) and a higher BW gain when their crypts were shorter. In conclusion, an hDDGS diet may facilitate small intestine longitudinal growth in broilers, which may subsequently improve dietary nutrient absorption. In addition, broiler chicks with shallow intestinal crypts exhibited better growth performance.

Plant‐based FRET biosensor discriminates environmental zinc levels

Heavy metal accumulation in the environment poses great risks to flora and fauna. However, monitoring sites prone to accumulation poses scale and economic challenges. In this study, we present and test a method for monitoring these sites using fluorescent resonance energy transfer (FRET) change in response to zinc (Zn) accumulation in plants as a proxy for environmental health. We modified a plant Zn transport protein by adding flanking fluorescent proteins (FPs) and deploying the construct into two different species. In Arabidopsis thaliana, FRET was monitored by a confocal microscope and had a 1.4-fold increase in intensity as the metal concentration increased. This led to a 16.7% overall error-rate when discriminating between a control (1μm Zn) and high (10mm Zn) treatment after 96h. The second host plant (Populus tremula×Populu salba) also had greater FRET values (1.3-fold increase) when exposed to the higher concentration of Zn, while overall error-rates were greater at 22.4%. These results indicate that as plants accumulate Zn, protein conformational changes occur in response to Zn causing differing interaction between FPs. This results in greater FRET values when exposed to greater amounts of Zn and monitored with appropriate light sources and filters. We also demonstrate how this construct can be moved into different host plants effectively including one tree species. This chimeric protein potentially offers a method for monitoring large areas of land for Zn accumulation, is transferable among species, and could be modified to monitor other specific heavy metals that pose environmental risks.

Planting Date Effect on Yield of Tomato, Eggplant, Pepper, Zinnia, and Snapdragon in High Tunnel in Mississippi

‘High tunnels’ are unheated greenhouse structures used to extend the growing season and protect high-value horticultural crops. High tunnels have been used for many years worldwide and their popularity has increased in Mississippi recently. A planting date study of ‘Roma’ tomato (Solanum lycopersicum L.), ‘Legend’ tomato, ‘Ichiban’ eggplant (Solanum melongena L.), ‘Sweet Banana’ pepper (Capsicum annuum L.), ‘Benarys Giant’ zinnia (Zinnia elegans L.), and ‘Potomac Red’ snapdragon (Antirrhinum majus L.) was conducted in 2010 in three high tunnels in Starkville, Mississippi. Each vegetable and cut flower cultivar was treated as an independent study. There were two planting dates for all the cultivars: 12 March 2010 and 2 April 2010. Only for zinnias, yield (272 stem/plot) of first planting date was higher than planting date two (106 stem/plot). A significant block effect was observed with ‘Legend’ tomato and ‘Ichiban’ eggplant where one high tunnel had significantly higher yield than the other two high tunnels. Harvesting of tomato, eggplant, and pepper from high tunnels was a month earlier than the field-grown crops. High tunnels can extend the growing season to provide produce to the market at earlier harvest dates in Starkville, Mississippi.