Delayed flowering has the potential to overcome the problem of restricted vegetative growth, prior to flowering, that is often associated with double-cropped soybeans [Grycine max (L.) Merr.]. Objectives were to study delayed flowering in soybeans as influenced by date of planting, to estimate the lengths of the component vegetative periods in soybeans under short-day conditions, and to study the mode of inheritance of delayed flowering in soybeans. Date of planting experiments conducted in the field at two Virginia locations using 27 cultivars and breeding lines showed that genotypic differences exist for delayed flowering, especially between delayed and normal flowering isolines. Lengths of the juvenile and inductive periods were estimated for some selected early and late flowering genotypes. F85-84l7 had a longer juvenile period, and F85-1226 had both longer juvenile and inductive periods than their respective early flowering isolines and cultivar Essex. cultivar. The method of moving plants from inductive short-days to long-days, which has been used to estimate the length of inductive period, was adapted to estimate the length of the juvenile period as well. Delayed flowering in soybeans appeared to be controlled by two loci, each with two alleles, and delayed flowering appeared to be recessive. Anyone of the genes in the homozygous recessive state delayed flowering. F85-1226 may be segregating for both genes while F85-84l7 appeared to contain only one. / Ph. D.

In-storage losses of seed vigor, i.e., aging, that occur between harvest and planting may leave soybeans essentially worthless as seed. Peroxidation of lipids, with resultant loss of membrane integrity, is theorized to be a primary event in seed aging. Lipid antioxidants might have the potential to protect dry seed by neutralizing free radicals, which propagate lipid peroxidation and other destructive events. Seed were treated with antioxidants in dimethyl sulfoxide (DMSO) to investigate possible protective effects of antioxidants during storage and thus also provide evidence for the lipid peroxidation model of seed aging. The toxicity of DMSO to soybean seed was found to be minimal at treatment times of 15 min or less. Seed were treated for 15 min with 0, 5, 25, and 50 mM solutions in DMSO of propyl gallate (PG), butylated hydroxyanisore (BHA), butylated hydroxy toluene (BHT), and tert-butylhydroquinone (TBHQ). Germination, seed leakage, seedling vigor, and phospholipids in the embryonic axis were monitored following storage at 40 C for up to 90 days. / Master of Science

Environmental stresses such as drought, salinity and low temperature have been major selective forces throughout plant evolution and are important factors which limit crop plant distribution and agricultural productivity. An understanding of how crops adapt to adverse conditions is not only of theoretical interest, but also has considerable practical value . Low-temperature stress subtraction libraries were constructed in a pBluescript vector with the two-step-PCR amplified cDNAs using subtractive hybridization. One insert cs18 was obtained and the sequence analysis of insert cs18 revealed that the insert cDNA had a 76% homology with the sequences of the corresponding portion of glucose dehydrogenase from Thermoplasma acidophilum and 62.0% homology with a genomic DNA of Arabidopsis. Four clones, cs18-13, cs18-14, cs18-15, and cs18-16 from low-temperature stress soybean root conventional cDNA library have been confirmed to have inserts that could hybridize to the cs18 insert. One cDNA with a Xba I and Xho I fragment of approximately 3,500 bp in length corresponded to the insert cs18 , which probably encodes for glucose dehydrogenase, was obtained. Northern blot analysis indicated that cs18 mRNA was highly expressed in soybean root but moderately expressed in leaves under low temperature. Changes in the nuclei of meristematic root cells in response to severe salinity were studied. Roots are in direct contact with the surrounding solution . Thus, they are the first to encounter the saline medium and are potentially the first site of damage or line of defence under salt stress. Nuclear deformation or degradation in the soybean root meristem with 150 mM or higher NaCI led to sequential cell degradation, cell death and cessation of plant growth . However, this study indicates that an increase in CaCI[2] concentration up to 5 mM could partially prevent salt injury to the cells. Tissue culture is an excellent tool for elucidat ing the correlation between plant organizational levels and salt tolerance because of the possibility it offers for studying the physiology of intact plantlets together with that of organs and single cells using homogenous plant material under uniform environmental conditions. One NaCI-tolerant cell line (R100) was isolated during this study. The R100 callus cell line was significantly more tolerant to salt than the salt-sensitive line (S100) during exposure to salt stress. Salt tolerance in this culture was characterized by an altered growth behaviour, reduced cell volume and relative water content, and accumulation of Na+, Cl ¯, K+, proline and sugars when grown under salt stress and with its subsequent relief. The selection of this salt tolerant cell line has potential for contributing new genetic variability to plant breeders. Sugars are not only important energy sources and structural components in plants , they are also central regulatory molecules controlling physiology, metabolism, cell cycle , development, and gene expression in plants. The concentrations of glucose and fructose increased during salt stress and after relieving salt stress, at a rate closely corresponding to the increase in relative water content. Their accumulation was the earliest response detected during the removing of salt stress indicating that glucose and fructose may play important roles during salt-stress. / Thesis (Ph.D.)-University of Natal, Pietermaritzburg, 2000.

A cytokinin habituated soybean callus has been isolated, utilizing the cytokinin soybean bioassay. The habituated callus line was subsequently characterized with a non-habituated callus line in relationship to levels of endogenous growth substances, ultrastructure, nitrogen metabolism and pattern of gene expression. The cytokinin habituated soybean callus contained a higher level of endogenous cytokinin-like activity in comparison to the non-habituated callus. This higher level of cytokinin present is in part due to a lower rate of degradation. The habituated callus tissue produced very low levels of ethylene, while the non-habituated callus produced ethylene at a much higher rate (57 fold higher), than the habituated callus. In contrast to what was found in habituated sugarbeet callus, only low levels of putrescine could be detected in both callus types. The putrescine content of habituated callus tissue was lower than that of non-habituated callus tissue. The ultrastructure of habituated callus cells exhibited several differences to what was observed in the non-habituated callus. Habituated callus cells appeared to have a thinner cell wall than that of the non-habituated callus cells. The cristae of the mitochondria in habituated cells were thicker than that of the non-habituated callus cells, indicating a lower metabolic activity. On day 14 of the growth period the nuclei of habituated callus demonstrated active RNA synthesis as indicated by the presence of several vacuolated nucleoli. Although no significant differences between proline levels of habituated callus and proline levels of non-habituated callus were observed, it was demonstrated that there was a difference in proline metabolism between the habituated and non-habituated calli. Utilizing an inhibitor of OAT, gabaculine, it was shown that in habituated callus tissue proline originated from ornithine during the first 14 days of growth. During the second half of the growth period, which characteristically consists of tissue with low biosynthetic activity, proline originated from glutamate. The production of proline in habituated callus from ornithine also corresponded to a period of high NH₄⁺ content in both callus types, while the production of proline from glutamate corresponded to a period of low NH₄⁺ content in the cells of both callus types. No such correlation was observed in proline metabolism of non-habituated callus. A similar turning point was observed in the activity of OAT of both callus types. Although the specific activity of OAT in both callus types mirrored their changes in RNA concentration, the percentage inhibition of OAT by gabaculine was not significant from day 14 in both callus types. This may indicate a change in the catalyzing properties of OAT in both callus types. It was further demonstrated that the non-habituated callus tissue contained some inhibitor inactivating OAT activity. With the use of gabaculine it was further shown that, in contrast to what was found in other habituated calli, there is no metabolic link between proline metabolism and putrescine synthesis. Both the habituated callus and the non-habituated callus exhibited a high nitrogen influx during the first 14 days of the growth period. The low NH₄⁺ content present in both callus types during the second half of the growth period coincided with higher levels of amino acids present in both callus types. The levels of precursor amino acids (glutamate, aspartate and alanine) did not fluctuate during the growth period, indicating a tight control on amino acid pools. Levels of amino acids further down the path of metabolism did not fluctuate drastically and there appeared to be very little difference between the levels of different amino acids measured in the habituated and nonhabituated calli. Serine was the dominant amino acid in both callus types. Total RNA concentrations of habituated callus were low in comparison to that of the non-habituated callus, except for a striking 12 fold increase on day 14 of the growth period. RNA concentrations of non-habituated callus increased gradually during the growth period and the highest concentration was recorded 21 days after subculturing. Several polypeptides were observed in the habituated callus that were not present in the non-habituated callus, utilizing IEF. Three polypeptides exhibited a change in concentration from day 6 to day 14 of the growth period in both the habituated and nonhabituated callus. These polypeptides appeared to decrease in nonhabituated callus, while they increased in the habituated callus. A complete cDNA library was constructed for both of the habituated and nonhabituated callus lines. Six different clones, that were over expressed in the habituated callus tissue, were isolated via subtractive techniques. One clone was characterized and showed homology to the glutamate/aspartate transport protein, the membrane component, of E. coli. / Thesis (Ph.D.)-University of Natal, Pietermaritzburg, 1998.

Thifensulfuron is a new herbicide of the sulfonylurea class under development by E. I. Dupont de Nemours Company Inc. for postemergence broadleaf weed control in soybeans [Glycine max (L.) Merr]. Field studies evaluated the influence of adjuvants and chlorimuron upon the efficacy of thifensulfuron. Thifensulfuron applied alone provided smooth pigweed (Amaranthus hybridus L. #AMACH) control at application rates 12% of those of the similar herbicide chlorimuron. Nonionic surfactant or crop oil concentrate increased soybean sensitivity to thifensulfuron, but an adjuvant was required to obtain consistent seedling common lambsquarters (Chenopodium album L. #CHEAL) control. Chlorimuron and thifensulfuron combinations did not control ivyleaf morningglory [Ipomoea hederacea (L.) Jacq. #IPOHE]. Greenhouse studies evaluated soybean cultivar sensitivity to thifensulfuron. Seven popular Virginia soybean varieties and one national variety (Williams 82) were screened for tolerance to thifensulfuron. Differences in varietal sensitivity was verified. Soybean varieties Vance, Essex, Hutcheson, and York proved to be more sensitive to 9.1 g ha⁻¹ thifensulfuron than FFR 561, Williams 82, or Deltapine 105. No relationship between sensitivity to thifensulfuron and Essex parentage could be drawn. The selectivity of the sulfonylurea class of herbicides is reportedly based on differential metabolism of the herbicide between sensitive and tolerant weed and crop species. Laboratory studies were conducted utilizing thifensulfuron-sensitive and tolerant weed species, velvetleaf (Abutilon theophrasti Medic. #ABUTH) and spurred anoda [Anoda cristata (L.) Schlecht #ANVCR], respectively, as well as the relatively tolerant Williams 82 and sensitive Vance soybean. Absorption and distribution studies indicated that all species absorbed and translocated similar amounts of ¹⁴C 1, 3, and 5 days after application of the methyl ester of [¹⁴C-thiophene] thifensulfuron. Metabolism studies indicated that both tolerant spurred anoda and sensitive velvetleaf metabolized thifensulfuron at similar rates 3 days after treatment. Metabolism appears to be the major mechanism for the selectivity of thifensulfuron to soybeans. The mechanism for spurred anoda tolerance to thifensulfuron has yet to be determined. This research indicates that broadcast foliar applications of 4.5 g ha⁻¹ thifensulfuron with 0.125% v/v nonionic surfactant or 1% v/v crop oil concentrate can provide selective postemergence smooth pigweed and common lambsquarters control for soybean production in Virginia. Caution should, however, be taken in prescribing greater than 4.5 g ha⁻¹ thifensulfuron due to the variability in cultivar sensitivity to thifensulfuron. / Ph. D.