Using the known sequences for serine palmitoyltransferase (SPT) and ceramide kinase (CERK) from Arabidopsis, candidates for the corresponding genes in Loblolly pine were cloned and examined during embryogenesis. The cloned two cDNA sequences from Loblolly pine, which has similarity of 81% and 88% respectively to two subunits of SPT1 and SPT2 in Arabidopsis, were presumed as the Loblolly pine SPT1 and SPT2 (Pt-SPT1 and Pt-SPT2). A few different versions of Pt-SPT1 mRNAs (2223 nts, 756 nts, 822 nts, and 754 nts respectively), most likely the alternative splicing results, were found. Three of these mRNAs are capable of encoding proteins. The long version (2223 nts) encodes a protein with 484 amino acids (Pt-SPT1); two short versions (822 nts, 756 nts) encode a 90 a.a. protein. Another cDNA sequence of 2396 nts encodes a protein of 493 a.a. (Pt-SPT2). Both predicted Pt-SPT1 and Pt-SPT2 proteins possess highly conserved serine palmitoyltransferase functional domains (E value 5.7e-61). Their expression patterns are different between somatic and zygotic embryogenesis. Two different versions of mRNAs, with 2786 nts (long), and 2320 nts (short) respectively, of ceramide kinases in Loblolly pine (Pt-CERKs) have been cloned. The long version encodes a protein with 721 a.a.; the short version with 560 a.a. The expression patterns for these two CERK mRNAs are different during embryo development. The long version is constitutively expressed, while the short one is only expressed in some stages with much lower expression level. Overexpression Pt-CERKL, Pt-CERKS, and Pt-CERKF in E.coli and function analysis in vitro show that all Pt-CERKs appear to have the same catalytic functions as their homologs in human and Arabidopsis, but with different efficiency. The catalytic efficiency was dramatically lower in the short Pt-CERK protein compared with the long Pt-CERK protein and Pt-CERKF. The membrane system is not necessary for the catalytic reactions of these three Pt-CERKs in vitro and Pt-CERKs were less dependent on the Ca2+ ions. Thus, these studies have provided the first information about SPT- and CERK- like proteins in loblolly pine, and open new avenues of investigation for the roles of sphingolipids in embryonic development.

myo-Inositol-1,2,3,4,5,6-hexakisphosphate (InsP6), also called phytic acid, is ubiquitous in eukaryotic cells and the most abundant inositol phosphate derivative. Loblolly pine (LP, Pinus taeda) constitutes the primary commercial species in the southern forest of U.S. Somatic embryogenesis (SE) is an effective technique to maintain the desirable genetic composition of the progeny and to accomplish the efficiency of propagation. SE can also serve as a tool for study of plant development. Unlike angiosperm embryos with attached cotyledons as seed storage organs, the diploid conifer embryo is surrounded by the unattached haploid female gametophyte (FG). In LP SE, FG tissue is absent in the embryogenic tissue culture. We found that extracts from early-stage FG stimulate growth and multiplication of early-stage somatic embryos, whereas FG water extracts from late stage contain substance(s) inhibitory to early-stage somatic embryo growth (DeSilva et al., 2007). We now present the isolation and identification of the inhibitory substance as InsP6 by means of water extraction, two gel filtrations and two ion exchange FPLC chromatographies. The results represent the first complete structural characterization of InsP6 from a natural product using LC/MS, LC/MS/MS, exact MS, 1D- and 2D-NMR analyses. We also report that there is a good correlation between the amount of InsP6 purified from FG tissue (1.3 nmoles per full-term FG) and the amount of InsP6 which inhibits somatic embryo growth. This novel approach of isolating and characterizing InsP6 from plant tissue, and investigating its role on SE can allow us to improve SE technology by circumventing current bottleneck, to elucidate enigmatic functions of InsP6 in plants, and most importantly, to utilize this molecule properly.

A system of loblolly pine growth and yield models was developed which used database files as input. Using database files as direct input created a compatible link between the growth and yield models and a Geographic Information System (GIS). Since growth and yield models can be used to update forest inventories and a GIS is a common method for maintaining forest inventory data, this compatibility provided a method to rapidly update past forest inventory records. This system was used to evaluate four different loblolly pine growth and yield models. The growth and yield models examined were: a whole stand, a diameter distribution - parameter prediction, a diameter distribution - parameter prediction, and an individual tree. Three different validation approaches were used to create fitting and validation data sets from permanent plot remeasurement data, and evaluate each of the four growth and yield models at varying projection periods. The periods used were zero, three, six, and nine years. Evaluations were based solely on the capability of each to model to predict merchantable volume. In terms of root mean square error of prediction, the individual tree and whole stand models performed superior than the diameter distribution models. At shorter projection periods the individual tree model performed better than the whole stand model, but the whole stand was superior at the nine year period. The parameter recovery models performed better for shorter periods than the parameter prediction model, but this difference diminished with longer periods. / Master of Science

This study explored the application of compost onto a small loblolly pine tree forest in northeast Texas. Its purpose was to determine if the application of various amounts of compost would provide for accelerated rates of growth for the trees. Soil parameters were also monitored. A total of 270 trees were planted and studied in a northeast Texas forest ecosystem. Compost rates of 5, 25, and 50 tons per acre with either soil or compost backfill were utilized and compared to a control without compost. Nonparametric and parametric ANOVA and Chi-Square tests were utilized. The results indicated that greater application rates retained greater moisture and higher pH levels in the soil. Compost applications also yielded a greater survival rate as well as larger tree height and diameter when compared to the control. The 25 ton/acre application backfilled in native soil achieved the greatest average in height and diameter when compared to the averages for the control plot. Greater growth differences for the 25S application can be attributed to additional nutrients coupled with a stable pH consistent with native soil acidity.

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