Improvement of staining method for observation of mitotic chromosomes in octoploid strawberry plants

An important strategy to conduct intentional breeding of octoploid strawberry plants is to recognize the functions of every chromosome. To do so, a methodology must be developed to distinguish chromosomes one by one. We reported the possibility of distinguishing chromosomes using light microscopy when somatic cells of octoploid strawberry plants were stained using ordinary methods with lacto-propionic orcein (LPO). However, karyotype analysis of octoploid strawberry plants required clearer chromosome images. This study obtained clearer chromosome images of octoploid Fragaria × ananassa and F. chiloensis plants. Three staining methods were examined: 60% acetic acid (AA) alone, 1.5% LPO alone, and two-step treatments with 60% AA and 1.5% LPO. Collected root tips of the plants were placed in 2 mM 8-hydroxyquinoline solution for 1 h and were subsequently stored at 4 °C for 15 h. The samples were then fixed in a 3:1 absolute alcohol: glacial acetic acid solution for 40 min, followed by mixture with 1N HCl solutions at room temperature for 2 h and then at 60 °C for 10 min. For separate staining using 60% AA and 1.5% LPO, the root tip was expelled on the glass slide with a drop of each solution for a few minutes to stain the chromosomes. For the two-step staining method, the samples stained with 60% AA were frozen at −80 °C for at least 5 min. The cover slip was removed using a razor blade. Subsequently, the specimens were air-dried and stained with the 1.5% LPO for 3 min. Digital images of chromosomes were obtained using light microscopy. Samples of the two-step staining method produced the clearest chromosome images in both F. × ananassa and F. chiloensis. Furthermore, the greatest color difference between the chromosomes and the cytoplasm was obtained from images of the two-step staining method among the three staining methods. These results demonstrate that the two-step staining method is useful for chromosome counting and karyotype analysis in strawberry plants.     

Analysis of phylogenetic relationships in Mangifera by restriction site analysis of an amplified region of cpDNA

The phylogenetic relationship of 13 Mangifera species collected in Thailand were examined. Among these species, M. foetida, M. odorata, and M. sylvatica have been cultivated as fruit crops for local markets in Thailand, as well as M. indica (common mango). The other nine wild species also seem to have potential values as rootstocks and breeding materials for tolerance to critical environmental conditions. By restriction fragment length polymorphism (RFLP) analysis of a particular region of cpDNA, these species were classified into two groups. The first group consisted of M. indica and M. sylvatica, and the second group consisted of M. caloneura, M. cochinchinensis, M. collina, M. flava, M. foetida, M. gedebe, M. griffithii, M. macrocarpa, M. oblongifolia, M. odorata, and M. pentandra. The species belonging to the same group are monomorphic for 20 restriction enzymes tested. The present study indicated low diversity in the amplified region of cpDNA among Mangifera species tested, even though the species in the second group were scattered to different subgenera and sections.     

Stem-rot damage and the progress of causal fungi in old-aged Japanese larch trees at the foot of Mt. Fuji

Damage caused by stem-rot and the progress of the causal fungi in old-aged Japanese larch (Larix kaempferi (Lamb.) Carr.) was investigated at the foot of Mt. Fuji. Stem-rot was found in 75% of 108 trees investigated, and volume of rot was 6% of the total wood volume in the forest investigated. Stem-rot damage was much greater than the damage by butt-rot.Stereum sanguinolentum (Alb. and Schw. ex Fr.) Fr. infected larch trees at the greatest incidence (49.4%). However,Porodaedalea chrysoloma (Fr.) Imaz. caused the most volume loss to the trees.S. sanguinolentum infected larch stems mainly through stem wounds, and decay caused by the fungus progressed 9.75×10² cm³/year on average.P. chrysoloma infected larch stems mainly through dead branches and wounds, and the average rate of decay progress for the fungus was 2.74×10³ cm³/year.     

Topographic variation in heterotrophic and autotrophic soil respiration in a tropical seasonal forest in Thailand

Soil respiration is a carbon flux that is indispensable for determining carbon balance despite variations over time and space in forest ecosystems. In Kanchanaburi, western Thailand, we measured the soil respiration rates at different slope positions—ridge (plot R), upper slope (plot U), and lower slope (plot L)—on a hill in a seasonal tropical forest [mixed deciduous forest (MDF)] to determine the seasonal and spatial variations in soil respiration on the slope. The heterotrophic (organic layer and soil) and autotrophic (root) respiration was differentiated by trenching. Soil respiration rates showed clear seasonal patterns: high and low rates in rainy and dry seasons respectively, at all plots, and tended to decrease up the slope. Soil respiration rates responded significantly to soil water content in the 0–30?cm layer, but the response patterns differed between the lower slope (plot L) and the upper slope (plots R and U): a linear model could be applied to the lower slope but exponential quadratic models to the upper slope. The annual carbon dioxide (CO₂) efflux from the forest floor was also associated with the slope position and ranged from 1908?gC?m^?2?year^?1 in plot L to 1199?gC?m^?2?year^?1 in plot R. With ascending position from plot L to R, the contribution of autotrophic respiration increased from 19.4 to 36.6% of total soil respiration, while that of the organic layer decreased from 26.2 to 9.4%. Mineral soil contributed to 46.3 to 54.4% of the total soil respiration. Soil water content was the key factor in controlling the soil respiration rate and the contribution of the respiration sources. However, the variable responses of soil respiration to soil water content create a complex distribution of soil respiration at the watershed scale.     

Effects of high temperature on flower colour and anthocyanin content in pink flower genotypes of greenhouse chrysanthemum (Chrysanthemum morifolium Ramat.)

Summary

The relationship between the intensity of flower colour and changes in the content of the main anthocyanins under various controlled temperatures was examined in order to clarify the effects of high temperature on flower colouration in six pink flower genotypes of greenhouse chrysanthemum (Chrysanthemum morifolium Ramat.). Poor colouration of flowers was observed at 30°C in all genotypes except ‘Chatoo’. This genotype showed little difference in flower colour between different temperature treatments. The degree of change in flower colour differed depending on the genotype, whereas no clear differences in flower colouring were observed between Summer – Autumn flowering and Autumn-flowering genotypes. All genotypes showed lower contents of the two anthocyanins tested [cyanidin 3-O-(6’’-O-monomalonyl- -glucopyranoside) and cyanidin 3-O-(3’’,6’’-O-dimalonyl- -glucopyranoside)] at higher temperatures. Therefore, flower colour changes were attributable to changes in these two main anthocyanins. Differences in colouration between genotypes and temperature conditions were also detectable in values that were measured using a colorimeter. Changed parameters that were visually verifiable were the a* value, representing the degree of red colour, and the C* value, representing chroma. For ‘Sei-Monako’, which showed visually greater differences between temperature treatments, the a* and C* values were low under high temperature conditions. On the other hand, in ‘Chatoo’, the differences detected by eye and those in a* and C* values between temperature treatments were small. In addition, the present results indicate that mean temperature is more important than either day or night temperature in determining the degree of flower colouration.     

Effective pollination period in durian (Durio zibethinus Murr.) and the factors regulating it

The effective pollination period (EPP) of durian was determined by both delayed and bud pollination, during which reproductive factors affecting the EPP, e.g., stigma receptivity, pollen tube growth in the style, and ovule longevity were studied histologically. This study was conducted in three distinct locations in Thailand, namely, the Chantaburi Horticultural Research Center and two private orchards in Chantaburi and Trat provinces. Results from artificial pollination revealed that at anthesis, the durian flower is receptive and has a high fruit set ratio. A mean fruit set of 50% was obtained at anthesis in the private orchard in Chantaburi province. However, the EPP of durian was found to be very short, lasting for only one night; the fruit set from pollination on the morning after anthesis ranged from 0% to 3.4%. No fruit set occurred following pollination 24 or more hours after anthesis. When compared with the flowers of other fruit species, the durian flower has a unique feature in that it blooms overnight; the following morning, there is abscission of all parts of the flower, except the gynoecium. Thus, EPP appears to be synchronized with flower longevity. On the other hand, the durian flower was receptive several hours before anthesis. The results of chemical tests, including the hydrogen peroxide test and Perex-Test^®, for the evaluation of stigma receptivity appeared to be in agreement with the EPP. However, fluorescent microscopy revealed that pollen could germinate even in the stigmas pollinated 48 h after anthesis, but the number of pollen tubes at the top of the style rapidly decreased from 34.6 (at anthesis) to 0.5 (48 h after anthesis). A correlation test demonstrated a higher correlation coefficient between the fruit set and the number of pollen than that between the fruit set and the result of Perex-Test^®. This indicated that pollen tube penetration or elongation in the style was inhibited, probably due to the deterioration of nutritional support from the pistil to the pollen tubes; this can be a limiting factor of the EPP in durian.     

Pathogenic Races of Phakopsora pachyrhizi on Soybean and Wild Host Plants Collected in Japan

A total of 45 single uredinial isolates of Phakopsora pachyrhizi were collected from rust-infected soybean and wild host plants (Pueraria lobata and G. soja ) at different localities in central and southwestern Japan. Eighteen pathogenic races were identified using a set of differential varieties composed of nine cultivars of soybean and two accession lines of G. soja. Nine and 11 races were found on soybean and wild host plants, respectively. Two races were common to soybean and wild host plants. Received 27 April 2001/ Accepted in revised form 22 August 2001