A comparison of leaf crystal macropatterns in the two sister genera Piper and Peperomia (Piperaceae)

Premise of the study: This is the first large-scale study comparing leaf crystal macropatterns of the species-rich sister genera Piper and Peperomia. It focuses on identifying types of calcium oxalate crystals and their macropatterns in leaves of both genera. The Piper results are placed in a phylogenetic context to show evolutionary patterns. This information will expand knowledge about crystals and provide specific examples to help study their form and function. One example is the first-time observation of Piper crystal sand tumbling in chlorenchyma vacuoles. Methods: Herbarium and fresh leaves were cleared of cytoplasmic content and examined with polarizing microscopy to identify types of crystals and their macropatterns. Selected hydrated herbarium and fresh leaf punches were processed for scanning electron microscopy and x-ray elemental analysis. Vibratome sections of living Piper and Peperomia leaves were observed for anatomical features and crystal movement. Key results: Both genera have different leaf anatomies. Piper displays four crystal types in chlorenchyma-crystal sand, raphides, styloids, and druses, whereas Peperomia displays three types-druses, raphides, and prisms. Because of different leaf anatomies and crystal types between the genera, macropatterns are completely different. Crystal macropattern evolution in both is characterized by increasing complexity, and both may use their crystals for light gathering and reflection for efficient photosynthesis under low-intensity light environments. Conclusions: Both genera have different leaf anatomies, types of crystals and crystal macropatterns. Based on Piper crystals associated with photosynthetic tissues and low-intensity light, further study of their function and association with surrounding chloroplasts is warranted, especially active crystal movement

T-URF13 Protein from Mitochondria of Texas Male-Sterile Maize (Zea mays L.)

The protein T-URF13 (URF13) is specific to mitochondria of maize (Zea mays L.) with Texas (T) male-sterile cytoplasm and has been implicated in causing male sterility and susceptibility to T-cytoplasm-specific fungal diseases. T-URF13 was purified from isolated mitochondria from maize (line B73) with T cytoplasm by gel filtration and a quasi two-dimensional polyacrylamide gel electrophoresis system. Antibodies to the purified and denatured protein were produced in rabbits. Anti-T-URF13 antiserum was used to show that T-URF13 is in the inner membrane of mitochondria and behaves as an integral membrane protein when mitochondria are fractionated with sodium carbonate or Triton X-114. The antiserum and protein A tagged with 20-nanometer-gold particles were used to localize T-URF13 in T mitochondria by electron microscopy of sections of isolated mitochondria from etiolated shoots and sections of roots and of tapetal cells at pre-and post-degeneration stages of microsporogenesis. The microscopic study confirms that T-URF13 is specifically localized in the mitochondrial membranes of all of the T mitochondria tested, notably those in the tapetum from the meiocyte stage to the late-microspore stage. No change in the amount of labeled T-URF13 protein in the mitochondria of aging tapetal cells was detected

Analysis and Mapping of Gene Families Encoding β-1,3-Glucanases of Soybean

Oligonucleotide primers designed for conserved sequences from coding regions of β-1,3-glucanase genes from different species were used to amplify related sequences from soybean [Glycine max (L.) Merr.]. Sequencing and cross-hybridization of amplification products indicated that at least 12 classes of β-1,3-glucanase genes exist in the soybean. Members of classes mapped to 34 loci on five different linkage groups using an F2 population of 56 individuals. β-1,3-Glucanase genes are clustered onto regions of five linkage groups. Data suggest that more closely related genes are clustered together on one linkage group or on duplicated regions of linkage groups. Northern blot analyses performed on total RNA from root, stem, leaf, pod, flower bud, and hypocotyl using DNA probes for the different classes of β-1,3-glucanase genes revealed that the mRNA levels of all classes were low in young leaves. SGlu2, SGlu4, SGlu7, and SGlu12 mRNA were highly accumulated in young roots and hypocotyls. SGlu7mRNA also accumulated in pods and flower buds

Soybean glucanases, compounds which encode therefor and related methods

The present invention provides 12 different sequences for soybean β-1,3-glucanases and the proteins for which they encode. Also provided are methods for the utilization of knockout mutants of the sequences which are useful for engineering genic male-sterile plants. Other methods and materials related to these sequences are also provided

The Genetics, Pathology, and Molecular Biology of T-Cytoplasm Male Sterility in Maize

This chapter reviews the genetics, pathology, and molecular biology of T-cytoplasm male sterility in maize. The chapter discusses the role of cytoplasmic male sterility systems in facilitating the production of hybrid seeds. The effects of widespread planting of T-cytoplasm maize on the severe 1970 epidemic and effect of a mitochondria1 gene on disease susceptibility and male sterility are discussed. It also discusses the involvement of nuclear cytoplasmic interactions in restoration of cms-T, the perspectives of cms-T researchers, and future directions. In cms-T plants, male sterility is associated with premature breakdown of the mitochondria-rich, tapetal cell layer of the anther; this layer is crucial to pollen production because it supplies nutrients to the developing microspores. In many species, cms is associated with the expression of novel open-reading frames in the mitochondrial genome. The studies provided a foundation for further research that resulted in the cloning of the T-urf13 and Rf2 genes from maize and the ChPKSl gene from C. heterostrophus, and the generation of models for the topology of urf13 in the inner mitochondrial membrane, Rfl-mediated processing of T-urfl3 transcripts, and the evolution of toxin biosynthesis in C. heterostrophus and M. zeae-maydis

Potato Cultivar Differences Associated with Mealiness

Russet Burbank, Norchip, Pontiac, and LaSoda potato cultivars were examined for the parameters mealy and waxy. Russet Burbank was judged dry, hard and particulate, typifying mealiness. Using phase contrast microscopy and scanning electron microscopy, raw mealy cells were determined to be larger and more irregularly shaped than cells from waxy cultivars. Mealy cooked cells were engorged with gelatinized starch, cell walls were more polarized, and cell shapes were better retained after mashing, when compared to waxy cells. NMR-T2 bound water readings from Russet Burbank and Pontiac samples did not differ from each other. Starch granule sizes and shapes varied by cultivar

The Genetics of Fertility in Soybean

Male and female reproductive structures play an important role in seed development in plants. Abnormalities in male or female reproductive structures can lead to sterility. In soybean, Glycine max (L.) Merr., about 75 sterility mutants have been identified and most of them have been mapped to chromosomes. Mapping results have shown that some chromosomal regions are hotspots for fertility genes. Fine mapping of some of the male-sterile, female-fertile mutants and male-sterile, femalesterile mutants resulted in identification of candidate genes for fertility. Sequence comparisons further helped in locating a few putative candidates. A CACTA- like transposable element that is responsible for reversion from sterility-to-fertility has been identified, and complete association between the presence of a transposon and sterility also has been shown. Several studies are underway that are using transformation sequences to clone fertility genes. Cloning and characterization of genes involved in male sterility and female sterility will help us recognize molecular mechanisms controlling sterility and help us understand the reproductive biology of soybean. This advancement of knowledge will assist in the development of a stable sterility system in soybean that can be utilized for hybrid seed production

Structural Adaptations in Overwintering Leaves of Thermonastic and Nonthermonastic Rhododendron Species

Evergreen rhododendrons (Rhododendron L.) are important woody landscape plants in many temperate zones. During winters, leaves of these plants frequently are exposed to a combination of cold temperatures, high radiation, and reduced photosynthetic activity, conditions that render them vulnerable to photooxidative damage. In addition, these plants are shallow-rooted and thus susceptible to leaf desiccation when soils are frozen. In this study, the potential adaptive significance of leaf morphology and anatomy in two contrasting Rhododendron species was investigated. R. catawbiense Michx. (native to eastern United States) exhibits thermonasty (leaf drooping and curling at subfreezing temperatures) and is more winter-hardy [leaf freezing tolerance (LT50) of containerized plants ≈–35 °C], whereas R. ponticum L. (native to central Asia) is less hardy (LT50 ≈–16 °C), and nonthermonastic. Thermonasty may function as a light and/or desiccation avoidance strategy in rhododendrons. Microscopic results revealed that R. ponticum has significantly thicker leaf blades but thinner cuticle than R. catawbiense. There is one layer of upper epidermis and three layers of palisade mesophyll in R. catawbiensecompared with two distinct layers of upper epidermis and two layers of palisade mesophyll in R. ponticum. We suggest that the additional layer of upper epidermis in R. ponticum and thicker cuticle and extra palisade layer inR. catawbiense represent structural adaptations for reducing light injury in leaves and could serve a photoprotective function in winter when leaf photochemistry is generally sluggish. Results also indicate that although stomatal density of R. ponticum is higher than that of R. catawbiense leaves, the overall opening of stomatal pores per unit leaf area (an integrated value of stomatal density and pore size) is higher by approximately twofold in R. catawbiense, suggesting that R. catawbiense may be more prone to winter desiccation and that thermonasty may be a particularly beneficial trait in this species by serving as a desiccation-avoidance strategy in addition to a photoprotection role

Microscopic Characterization of a Transposon-Induced Male-Sterile, Female-Sterile Mutant in Glycine max L.

Premise of research. A male-sterile, female-sterile mutant was discovered in a w4-m mutable line of Glycine max L. The mechanism of its sterility was not well understood. Therefore, different cytological and microscopic techniques were undertaken to better understand the process of mutant phenotype development. Molecular research indicated that mer3 was responsible for the sterility. Methodology. Macro images were collected of whole plants, flowers, anthers, pods, and ovules. Chromosome spreads from anthers at various meiotic stages were examined. Confocal scanning laser microscopy using optical sectioning was utilized on whole anthers and ovules at various developmental stages. Whole mature anthers and isolated pollen images were collected and studied with SEM. Pivotal results. In observations of the mutant, male cell development was found to begin normally and then digresses at metaphase I of meiosis, when abnormal segregation of chromosomes with reduced bivalent formation was observed. It was the abnormal formation of univalents and bivalents that led to male sterility. On the female side, the progression of development was arrested in the megagametophyte stage likely because of abnormal meiosis, leading to ovule abortion and female sterility. Conclusions. The G. max male-sterile, female-sterile mutant was shown to have the same phenotype of mer3 sterility already shown in Arabidopsis, rice, yeast, and some animal systems

A microscopic study of the trichomes on gynoecia of normal and tetraploid Clark cultivars of Glycine max and seven near isogenic lines

The surfaces of gynoecia of Glycine max cultivars—normal Clark, a tetraploid Clark, and seven isolines—display variations in at least three types of trichomes. The normal Clark soybean gynoecium has at least three and possibly four types of trichomes: a two‐ to four‐celled, elongate, thick‐walled trichome (TWT), an elongate thin‐walled unicellular trichome (UCT), a secretory multicellular trichome (MCT), and an elongate thin‐walled bicellular trichome that we have interpreted as an immature TWT. All these types are present on the gynoecium by 1 d before anthesis. After fertilization, the UCT is rare, but the other types continue to initiate and develop on the young pod. During flowering, the UCT and the MCT are distributed along the ovary from the base of the gynoecium to the top of the ovary. The TWT forms primarily along the dorsal side of the style. The Clark tetraploid and the extra-dense, dense 1, dense 2, sharp hair tip, sparse, and puberulent isolines have all four types of trichomes on their gynoecia, although TWTs and UCTs are very short in the puberulent isoline. The glabrous isoline is missing the TWT but has a short, thin‐walled trichome that occurs infrequently, mainly along the dorsal side of the style. The ratio of TWTs on the gynoecium of the normal Clark to those on the isolines is 1:4 on extra-dense, 1:3 on dense 2, 1:1.6 on dense 1, 1:2 on the tetraploid, 1:0.5 on the sparse, and 1:0.2 on the puberulent. There is a positive correlation between the expanded distribution of TWTs onto other parts of the gynoecium and increased pubescence of TWTs on the gynoecium. The numbers and morphology of MCTs are similar in the normal Clark, the tetraploid, and the seven isolines, except for extra-dense, where the numbers of MCTs are suppressed. The numbers of UCTs were significantly greater on the extra-dense, dense 2, and dense 1 isolines than on the normal Clark, but distribution was the same. During early pod development, TWTs and MCTs are most prominent, indicating that the UCTs either disappear or may be a precursor of TWTs. No functions are known for any of the types of trichomes on soybean gynoecia, but possible roles are discussed