Chromosomal location of genes controlling flavonoid production in hexaploid wheat

Two-dimensional paper chromatography was performed on methanol extracts of leaves of hexaploid bread wheat, Triticum aestivum L. em. Thell. cultivar Chinese Spring, and of the available nullisomic-tetrasomic compensating lines, the tetrasomic lines and the ditelocentric lines. The chromatograms had 27 spots identified as flavonoids and six representing phenolic acids. Some of the areas were complex and contained more than one compound. Four flavonoids were identified as under the control of gene(s) on chromosome arms 1DS, 4DL, 5AS and 6BS. A phenolic glycoside was concluded to be controlled by a gene(s) on chromosome arm 7BL. Gene(s) on chromosome arm 4DL affected the amount of compounds in two other spots, and gene(s) on chromosome arm 4BS reduced the level of all flavonoid compounds. The individual compounds in some of the complex spots may be under the control of gene(s) on homoeologous chromosomes.     

Cytoplasmic filaments and cellular wound healing in amoeba proteus

The flexibility and self-healing properties of animal cell surface membranes are well known. These properties have been best exploited in various micrurgical studies on living cells (2, 3), especially in amoebae (7, 20). During nuclear transplantation in amoebae, the hole in the membrane through which a nucleus passes can have a diameter of 20-30 μm, and yet such holes are quickly sealed, although some cytoplasm usually escapes during the transfer. While enucleating amoebae in previous studies, we found that if a very small portion of a nucleus was pushed through the membrane and exposed to the external medium, the amoeba expelled such a nucleus on its own accord. When this happened, a new membrane appeared to form around the embedded portion of the nucleus and no visible loss of cytoplasm occurred during nuclear extrusion. In the present study, we examined amoebae that were at different stages of expelling partially exposed nuclei, to follow the sequence of events during the apparent new membrane formation. Unexpectedly, we found that a new membrane is not formed around the nucleus from inside but a hole is sealed primarily by a constriction of the existing membrane, and that cytoplasmic filaments are responsible for the prevention of the loss of cytoplasm.     

Saikosaponin-d,a novel SERCA inhibitor,induces autophagic cell death in apoptosis-defective cells

Autophagy is an important cellular process that controls cells in a normal homeostatic state by recycling nutrients to maintain cellular energy levels for cell survival via the turnover of proteins and damaged organelles. However, persistent activation of autophagy can lead to excessive depletion of cellular organelles and essential proteins, leading to caspase-independent autophagic cell death. As such, inducing cell death through this autophagic mechanism could be an alternative approach to the treatment of cancers. Recently, we have identified a novel autophagic inducer, saikosaponin-d (Ssd), from a medicinal plant that induces autophagy in various types of cancer cells through the formation of autophagosomes as measured by GFP-LC3 puncta formation. By computational virtual docking analysis, biochemical assays and advanced live-cell imaging techniques, Ssd was shown to increase cytosolic calcium level via direct inhibition of sarcoplasmic/endoplasmic reticulum Ca²⁺ ATPase pump, leading to autophagy induction through the activation of the Ca²⁺/calmodulin-dependent kinase kinase–AMP-activated protein kinase–mammalian target of rapamycin pathway. In addition, Ssd treatment causes the disruption of calcium homeostasis, which induces endoplasmic reticulum stress as well as the unfolded protein responses pathway. Ssd also proved to be a potent cytotoxic agent in apoptosis-defective or apoptosis-resistant mouse embryonic fibroblast cells, which either lack caspases 3, 7 or 8 or had the Bax-Bak double knockout. These results provide a detailed understanding of the mechanism of action of Ssd, as a novel autophagic inducer, which has the potential of being developed into an anti-cancer agent for targeting apoptosis-resistant cancer cells.     

Evolutionary implications of substitution patterns in prolamin genes of Oryza glaberrima (African rice, Poaceae) and related species

Patterns of sequence variation of nuclear genes encoding 10-kDa and 16-kDa prolamin seed storage proteins were examined in Oryza glaberrima (African rice, Poaceae) and O. barthii and compared to available sequences for the genus to assess potential application of these gene families in evolutionary studies. Sequence variation among species in 10-kDa genes was very low. In contrast, the 16-kDa genes have undergone rapid evolution, displaying a larger number of length and point mutations that in some cases result in frame shift or produce truncated protein or pseudogenes. The proportion of nonsynonymous substitution is high in both genes. Although nonsynonymous mutations did not alter the overall profile of the protein, pronounced shifts in proportions of some amino acids were evident and could have systematic application. The data provide support for a proposed direct evolution of the Asian (O. sativa) and African rice from O. rufipogon and O. barthii, respectively. Patterns of amino acid frequencies of the 10-kDa genes show the distinctness of O. rufipogon and O. longistaminata from the other species. The study underscores the potential application of the prolamin genes as markers from the nuclear genome for evolutionary studies in grasses at different taxonomic levels.     

Characterization of 10 KDA prolamin genes in Phyllostachys aurea (Bambusoideae, Poaceae)

Prolamin is the dominant class of seed storage protein in grasses (Poaceae). Information on the 10 kDa multigene family coding for prolamins characteristic of the bambusoid-oryzoid grasses is limited. Two genes encoding 10 kDa prolamin were cloned and sequenced in the bambusoid species Phyllostachys aurea to assess the sequence diversity of this gene family in the oryzoid-bambusoid grasses. The genes, ~417 bp in length, were 96% similar at the DNA sequence level, differing in 12 base substitutions dispersed throughout the sequence. Eight of these mutations were nonsynonymous, leading to amino acid substitutions in the coding region, and one was nonsense, producing an amber stop codon. One gene had an open reading frame (ORF) of 139 amino acids, while the other gene had a shorter ORF (106 amino acids) due to the presence of a stop codon in the coding region and, thus, represents a pseudogene. Deduced proteins showed amino acid composition similar to that of rice. The study underscores the overall conserved nature of this multigene family and reflects considerable sequence divergence at the DNA and amino acid levels between the Oryza and the Phyllostachys genes. The systematic implication of the data is discussed in light of the inconsistent placement of Oryza in the Bambusoideae or Oryzoideae.     

CLUSTER ANALYSIS OF ANTHER-DERIVED PLANTS OF SOLANUM PHUREJA (SOLANACEAE) BASED ON MORPHOLOGICAL CHARACTERISTICS

The feasibility of grouping anther-derived plants of Solarium phureja according to ploidy based on their morphological characteristics was studied. Canonical discriminant analysis identified four characteristics (anther length, number of chloroplasts per pair of guard cells, leaf width, corolla width at widest diam) of nine measured as the most effective combination for diagnosing ploidy. Data for these characteristics from two sets of plants were subjected to two clustering techniques, one using the average linkage clustering (UPGMA of the NT-SYS programs) and the other using centroid sorting (SAS-Fastclus). Screening of anther-derived plants by cluster analysis proved to be an efficient means of separating monoploids from the other ploidy levels.     

Impact of missing data,gene choice,and taxon sampling on phylogenetic reconstruction: the Caryophyllales (angiosperms)

Density of taxon sampling and number/kind of characters are central to achieving the ultimate goals in phylogenetic reconstruction: tree robustness and improved accuracy. In molecular phylogenetics, DNA sequence repositories such as GenBank are potential sources for expanding datasets in two dimensions, taxa and characters, to the level of “supermatrices.” However, the issue of missing characters/genomic regions is generally considered a major impediment to this endeavor. We used here the angiosperm order Caryophyllales to systematically address the impact of missing data when expanding taxon sampling and number of characters in phylogenetic reconstruction. Our analyses show that expansion of taxon sampling by ~13-fold resulted in improved phylogenetic assessment of the Caryophyllales despite up to 38% missing data. Expanding number of characters in the dataset by allowing for up to 100-fold increase in amount of missing data and inclusion of entries with about 40% missing genomic regions did not negatively impact tree structure or robustness, but to the contrary improved both. These results are timely regarding the ongoing efforts to achieve detailed assessment of the tree of life.     

RACIAL EVOLUTION IN ELEUSINE CORACANA SSP. CORACANA (FINGER MILLET)

Eleusine coracana ssp. coracana (finger millet, eleusine) is widely distributed in Africa and India as a cereal. The crop is cultivated in diverse eco-geographical areas. Over this range of distribution, eleusine displays high variability in vegetative, floral and seed morphology. The intent of this study is to establish correlations between morphology and distribution, and to determine the intraspecific taxonomy of the crop. Three eco-geographical races were determined: (1) an African highland race which is cultivated in the East African highlands, (2) a lowland race which is grown in the lowlands of Africa and South India, and (3) an Indian race with its center of distribution in Northeast India. In addition to these basic races, an Indian highland type was identified. The African highland race is the most primitive from which the lowland race evolved. The latter race was subsequently introduced to southern India where a secondary center of diversity became established. The Indian race originated from the lowland race, while the North Indian highland type can be derived from either or both of the two basic races in India. This study indicated that natural selection played a major role in the evolution of the crop. Artificial selection, although significant, was restricted within the limits imposed on it by the ultimate adaptation of the races to their environments.     

ORIGIN OF TRIPSACUM ANDERSONII (GRAMINEAE)

Tripsacum andersonii Gray (Gramineae) is a species with 2n = 64 chromosomes. Chromosome behaviour during meiosis of microsporogenesis suggests that the species combines three homologous haploid Tripsacum genomes of x = 18 (54 chromosomes), and an alien haploid genome of x = 10 chromosomes. Cytogenetic studies indicate that T. andersonii originated as a hybrid between a species of Tripsacum (2n = 36) and a species of Zea (2n = 20). Comparative morphology and flavonoid chemistry fail to identify the Zea species involved in this intergeneric hybrid. Chromosome morphology suggests that it was either Z. mays L. subsp. mays (domesticated maize) or subspecies mexicana (Schrad.) Iltis (annual teosinte). The Tripsacum parent probably was T. latifolium Hitchc. of Central America. It resembles T. andersonii in vegetative morphology. Tripsacum maizar Hernandez et Randolph and T. laxum Nash, which resemble T. andersonii in flavonoid chemistry, are eliminated as possible parents on the basis of growth habit and the morphology of their hybrids with maize.