Robert J. Jones

Low water potential disrupts carbohydrate metabolism in maize (Zea mays L.) ovaries

Water deficit during pollination increases the frequency of kernel abortion in maize (Zea mays L.). Much of the kernel loss is attributable to lack of current photosynthate, but a large number of kernels fail to develop on water-deficient plants even when assimilate supply is increased. We examined the possibility that assimilate utilization by developing ovaries might be impaired at low water potential ([Psi]w). Plants were grown in the greenhouse in 20-L pots containing 22 kg of amended soil. Water was withheld on the first day silks emerged, and plants were hand-pollinated 4 d later when leaf [Psi]w decreased to approximately - 1.8 MPa and silk [Psi]w was approximately -1.0 MPa. Plants were rehydrated 2 d after pollination. The brief water deficit inhibited ovary growth (dry matter accumulation) and decreased kernel number per ear by 60%, compared to controls. Inhibition of ovary growth was associated with a decrease in the level of reducing sugars, depletion of starch, a 75-fold increase in sucrose concentration (dry weight basis), and inhibition of acid invertase (EC 3.2.1.26) activity. These results indicate that water deficits during pollination disrupt carbohydrate metabolism in maize ovaries. They suggest that acid invertase activity is important for establishing and maintaining reproductive sink strength during pollination and early kernel development.

Cytokinin Oxidase Gene Expression in Maize Is Localized to the Vasculature, and Is Induced by Cytokinins, Abscisic Acid, and Abiotic Stress

Cytokinins are hormones that play an essential role in plant growth and development. The irreversible degradation of cytokinins, catalyzed by cytokinin oxidase, is an important mechanism by which plants modulate their cytokinin levels. Cytokinin oxidase has been well characterized biochemically, but its regulation at the molecular level is not well understood. We isolated a cytokinin oxidase open reading frame from maize (Zea mays), called Ckx1, and we used it as a probe in northern and in situ hybridization experiments. We found that the gene is expressed in a developmental manner in the kernel, which correlates with cytokinin levels and cytokinin oxidase activity. In situ hybridization with Ckx1 and transgenic expression of a transcriptional fusion of the Ckx1 promoter to the Escherichia coli β-glucuronidase reporter gene revealed that the gene is expressed in the vascular bundles of kernels, seedling roots, and coleoptiles. We show that Ckx1 gene expression is inducible in various organs by synthetic and natural cytokinins. Ckx1 is also induced by abscisic acid, which may control cytokinin oxidase expression in the kernel under abiotic stress. We hypothesize that under non-stress conditions, cytokinin oxidase in maize plays a role in controlling growth and development via regulation of cytokinin levels transiting in the xylem. In addition, we suggest that under environmental stress conditions, cytokinin oxidase gene induction by abscisic acid results in aberrant degradation of cytokinins therefore impairing normal development.

Minireview: nucleotide prodrugs

Nucleotides have shown interesting biological activities in a wide variety of antiviral, antiproliferative, immunomodulatory and other biological assays, and they present promising drug candidates. Because of their negative charge(s) nucleotides suffer from some disadvantages which can be successfully overcome by the utilization of nucleotide prodrugs. Nucleotide prodrugs were successfully used to increase oral absorption of nucleotides in vivo. By taking advantage of intracellular triggers (reducing potential, enzyme activity, pH), nucleotide prodrugs can be used in vitro for the intracellular delivery of the nucleotide resulting in enhanced potency and in some cases enhanced selectivity. Nucleotide prodrugs have also been utilized for tissue specific delivery of the nucleotides in vivo resulting in altered selectivity and reduced toxicity. For nucleotide prodrugs, their ultimate intended use is (in most cases) in vivo for the treatment of a disease. Thus, it is important to incorporate adequate assays and design criteria into any prodrug effort. In vivo systems are complicated because of metabolism, excretion and tissue distribution of the prodrug and the parent. Thus, results of in vitro assays have to be interpreted cautiously because they may be unsuitable predictors of the in vivo situation.

Disruption of Maize Kernel Growth and Development by Heat Stress (Role of Cytokinin/Abscisic Acid Balance)

Temperature stress during kernel development affects maize (Zea mays L.) grain growth and yield stability. Maize kernels (hybrid A619 x W64A) were cultured in vitro at 3 d after pollination and either maintained at 25[deg]C or transferred to 35[deg]C for 4 or 8 d, then returned to 25[deg]C until physiological maturity. Kernel fresh and dry matter accumulation was severely disrupted by the long-term heat stress (8 d at 35[deg]C) and did not recover when transferred back to 25[deg]C, resulting in abortion of 97% of the kernels. Kernels exposed to 35[deg]C for 4 d (short-term heat stress) exhibited a recovery in kernel growth and water content at about 18 d after pollination and kernel abortion was reduced to about 23%. During the cell division phase, abscisic acid (ABA) levels showed a steady decline in the control but maintained a moderate level in the heat-stressed kernels. However, later in development heat-stressed kernels had significantly higher levels of ABA than the control. Cytokinin analysis confirmed a peak in zeatin riboside and zeatin levels in control kernels at 10 to 12d after pollination. In contrast, kernels subjected to 4 d of heat stress had no detectable levels of zeatin and the zeatin riboside peak was reduced by 70% and delayed until 18 d after pollination. The long-term heat-stressed kernels showed low to nondetectable levels of either zeatin riboside or zeatin. Regression analysis of ABA level against cytokinin level during the endosperm cell division phase revealed a highly significant negative correlation in nonstressed kernels but no correlation in kernels exposed to short-term or long-term heat stress. Application of benzyladenine to heat-stressed, growth-chamber-grown plants increased thermotolerance in part by reducing kernel abortion at the tip and middle positions on the ear. These results confirm that shift in hormone balance of kernels is one mechanism by which heat stress disrupts maize kernel development. The maintenance of high levels of cytokinins in the kernels during heat stress appears to be important in increasing thermotolerance and providing yield stability of maize.

Role and function of cytokinin oxidase in plants

Cytokinin oxidase (CK oxidase) is widely distributed in plants and is the only enzyme that has been shown unequivocally to catalyze the catabolism of specific cytokinins (CKs) to inactive products that lack the N6-unsaturated side chain. Thus, the enzyme is thought to play a major role in controlling the level or species of CKs in plant tissues. However, despite its discovery more than 25 years ago, little attention has been given to the elucidation of its role and function in plant growth and development. This review seeks to bring in to context the current state of knowledge regarding the biochemical and molecular properties, regulation in undifferentiated and differentiated tissues, and recent results from studies using transgenic plants in an attempt to provide a more comprehensive understanding of the physiological significance of the enzyme in plants. Notwithstanding species, tissue and other specific differences, in general, CK oxidase appears to contribute to CK homeostasis in plants. However, complete clarity as to its function awaits purification of the protein to homogeneity and the ultimate development of requisite molecular probes.

Magnetic Resonance Imaging of the Idiopathic Inflammatory Myopathies : Structural and Clinical Aspects

Idiopathic inflammatory myopathies are chronic diseases clinically characterized by symmetrical proximal muscle weakness. MRI has assumed a major role in the evaluation and management of these conditions. It is sensitive to the presence of inflammation and edema, especially with incorporation of fat suppression sequences, so it is a useful tool for establishing an early diagnosis, for evaluating the extent and number of lesions, and for determining the right site for biopsy. The noninvasive nature of the procedure makes it ideal for serial studies to evaluate response to treatment. Whole‐body MRI can scan a large volume of muscles without prolonged acquisition time and has the potential to identify previously unsuspected sites of involvement. MRI is also an excellent technique for identifying areas of fatty infiltration within the muscles, which usually occurs in the late stages of inflammatory myopathies. In summary, MRI has revolutionized the way muscular diseases are diagnosed and treated.

Development of Antisense Therapeutics: Implications for Cancer Gene Therapy

Gene inhibition by antisense oligodeoxynucleotides (ODNs) has been documented for numerous oncogenes important in human cancers.*.2 Many of the early studies led to speculation on the use of antisense ODNs in therapies. Unfortunately, more recent studies appear to indicate that much of the activity attributed to an antisense mechanism is due to a nonspecific and non-antisense mechanism of Advances in the antisense field will require new modifications that enhance cellular permeation, affinity, and stability of ODNs.

Oligodeoxynucleotides containing 5-(1-propynyl)-2'-deoxyuridine formacetal and thioformacetal dimer synthons

Abstract Oligodeoxynucleotides containing the C-5 propyne 2′-deoxyuridine analog in conjunction with the formacetal and 3′-thioformacetal linkage are described. Thermal denaturation analysis demonstrates that these analogs have enhanced binding affinity to both single-strand RNA and DNA and double-strand DNA.

In vitro Culture of Maize Kernels

This method provides a means with which to study the development of intact maize kernels under conditions in which nutritional, hormonal, environmental and other factors can be modified and controlled. Kernels may be cultured from 2 days after pollination up to physiological maturity. Final kernel size may vary, but average kernel dry weight generally ranges from 70% to 100% of normal.

Effect of 1-aminocyclopropane-1-carboxylic acid on maize kernel development in vitro.

Pollination stimulates ethylene production in maize ears, and the application of ethephon during the pollination period can cause kernel abortion. The objective of this study was to determine if kernel abortion could be induced in vitro by the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC). Adding ACC to the culture medium resulted in the evolution of ethylene which caused abortion and reduced mature kernel mass. The effect of ethylene on kernel abortion and dry matter accumulation was partially negated by the addition of the ethylene-binding site inhibitor, 2,5-norbornadiene (NBD). The effect of ethylene on kernel abortion was greatest during the early stage of kernel development and was intensified by an increase in media sucrose concentration. These data suggest that ethylene could regulate kernel abortion in maize.