Taxonomic status of Monotropastrum humile, with special reference to M. humile var. glaberrimum (Ericaceae, Monotropoideae).

Taxonomic treatment of the achlorophyllous monotropoid plant Monotropastrum humile is still unclear and confusing because of the lack of detailed morphological analyses and molecular phylogeny. In particular, the taxonomic status of a glabrous variety, M. humile var. glaberrimum, is under debate. Our detailed examination of the morphological characteristics of living plants revealed that M. humile var. glaberrimum can be easily distinguished from the putative conspecific taxon M. humile var. humile by characteristics not previously recognized, namely the shape and color of the floral disc. Most morphological features characterizing Cheilotheca were also found in M. humile var. glaberrimum. Moreover, there was considerable nucleotide differentiation in the internal transcribed spacer (ITS)2 sequences of M. humile var. humile and var. glaberrimum. Molecular analysis of the phylogenetic relationship of M. humile var. humile, var. glaberrimum, and other monotropoids using ITS2 sequences showed that two varieties of M. humile formed a monophyletic clade with a member of a different genus, Monotropa L., but obvious phylogenetic relationships among these three taxa were not obtained. Thus we conclude that Monotropastrum humile var. glaberrimum should be treated as a distinct species. However, the generic affiliation of M. humile var. glaberrimum could not be determined because of its intermediate character state combination and the insufficient characterization of related species. We strongly suggest that Monotropastrum as a whole needs re-evaluation.

Insight into the basis of root growth in Arabidopsis thaliana provided by a simple mathematical model.

Plant organ growth changes under genetic and environmental influences can be observed as altered cell proliferation and volume growth. The two aspects are mutually dependent and intricately related. For comprehensive growth analysis, it is necessary to specify the relationship quantitatively. Here, we develop a simple mathematical model for this purpose. Our model assumes that the biological activity of a given organ is proportional to the cell number of the organ and is allocated into three aspects: cell proliferation, volume growth, and organ maintenance. We analyzed the growth of primary roots of Arabidopsis thaliana (L.) Heynh. in one tetraploid and four diploid strains using this model. The analysis determined various growth parameters, such as specific cost coefficients of cell proliferation and volume growth for each strain. The results provide insight into the basis of interstrain variations and ploidy effects in root growth.

Large-scale general collection of wild-plant DNA in Mustang, Nepal.

The deposit of DNA samples of wild plants that correspond to voucher specimens is highly informative and greatly enhances the value of the herbarium specimens. The Society of Himalayan Botany (SHB), Tokyo, has assembled general collections of flowering plants of the Sino-Himalayan region for more than 40 years. In a trial of the collection of these types of bioresources for use in basic research, we adopted FTA cards, which have recently been used for large-scale collection of DNA of humans, microorganisms and viruses, for the general collection of DNA samples of wild plants during a botanical expedition in Mustang, Nepal, in 2003. Three hundred and fifty-five plant specimens from Mustang, Nepal, were collected along with the corresponding DNA samples. Examination of the quality of the DNA samples by PCR demonstrated the utility of the collection system. The identification of all of the specimens collected, as well as data from the specimens, will be presented on the Flora of Nepal Database website (http://ti.um.u-tokyo.ac.jp/default.htm), which is open to the public. The DNA resources will be identified on the website and distributed openly by the SHB to researchers worldwide for basic research.

Development and structure of trichotomous branching in Edgeworthia chrysantha (Thymelaeaceae).

We studied the development and structure of the unusual trichotomous branching of Edgeworthia chrysantha. Three "branch primordia" are formed sequentially on the shoot apex of a main axis and develop into trichotomous branching. The branch primordia are clearly distinguishable from the typical axillary buds of other angiosperms; they develop much more rapidly than axillary buds, and the borders between the branch primordia and shoot apex of the main axis are anatomically unclear. Furthermore, at a later stage, leaves subtending the branch primordia produce typical axillary buds. These results suggest that the trichotomous branching in this species involves the division of the shoot apical meristem. Expression analysis of genes involved in branching or maintenance of the shoot apical meristem in this species should clarify the control mechanism of this novel branching pattern in angiosperms. We also observed the phyllotactic patterns in trichotomous branching and have related these patterns to the shoot system as a whole.

Flavonoids in translucent bracts of the Himalayan Rheum nobile (Polygonaceae) as ultraviolet shields.

UV-absorbing substances were isolated from the translucent bracts of Rheum nobile, which grows in the alpine zone of the eastern Himalayas. Nine kinds of the UV-absorbing substances were found by high performance liquid chromatography (HPLC) and paper chromatography (PC) surveys. All of the five major compounds are flavonoids, and were identified as quercetin 3- O-glucoside, quercetin 3- O-galactoside, quercetin 3- O-rutinoside, quercetin 3- O-arabinoside and quercetin 3- O-[6"-(3-hydroxy-3-methylglutaroyl)-glucoside] by UV, 1H and 13C NMR, mass spectra, and acid hydrolysis of the original glycosides, and direct PC and HPLC comparisons with authentic specimens. The four minor compounds were characterised as quercetin itself, quercetin 7- O-glycoside, kaempferol glycoside and feruloyl ester. Of those compounds, quercetin 3- O-[6"-(3-hydroxy-3-methylglutaroyl)-glucoside] was found in nature for the first time. The translucent bracts of R. nobile accumulate a substantial quantity of flavonoids (3.3-5 mg per g dry material for the major compounds). Moreover, it was clarified by quantitative HPLC survey that much more of the UV-absorbing substances is present in the bracts than in rosulate leaves. Although the flavonoid compounds have been presumed to be the important UV shields in higher plants, there has been little characterisation of these compounds. In this paper, the UV-absorbing substances of the Himalayan R. nobile were characterised as flavonol glycosides based on quercetin.

Floral development and phyllotactic variation in Ceratophyllum demersum (Ceratophyllaceae).

The floral development of staminate and pistillate flowers of Ceratophyllum demersum was observed, with particular focus on the phyllotactic variation in staminate flowers, using scanning electronic microscopy (SEM). We discerned patterns of development of some important new morphological features, e.g., the difference and discontinuity between the organ initiation in stamens and that in bracts (or tepals) and the initial presence of a mucilaginous appendage on each pistil. Female flowers are considered to be very specialized through reduction. In male flowers stamen initiation changes between early and late floral development. The four or five stamens in the outermost whorl initiate first on the abaxial and lateral sides of the floral apex and only later on the adaxial side (unidirectional). Later the inner stamens initiate spirally, and this is the main pattern in the stamen initiation. Members of each whorl differ among themselves in time of initiation and in ultimate size. The phyllotactic variation in staminate flowers of Ceratophyllum, suggested by previous studies, is derived from the variation in stamen number and the difference of stamen initiation between the early and later stages. The development in Ceratophyllum has some similarities to those of ANITA plants except for Nymphaeales.

Photosynthetic characteristics of a giant alpine plant, Rheum nobile Hook. f. et Thoms. and of some other alpine species measured at 4300 m, in the Eastern Himalaya, Nepal.

The photosynthetic characteristics of a giant alpine plant, Rheum nobile Hook. f. et Thoms. and of some other alpine species were studied in situ at 4300 m, in the Eastern Himalaya, Nepal, during the summer monsoon season. Although rainy and overcast weather was predominant, the daytime photon flux density (400-700 nm) ranged from 300 to 500 µmol quanta m-2 s-1. Under such conditions, the temperature of leaves of R. nobile ranged from 10 to 14°C, and the rate of photosynthetic CO2 exchange ranged from 10 to 16 µmol CO2 m-2 s-1. The ratios of the maximum rate of photosynthetic CO2 fixation to leaf nitrogen content (defined as instantaneous nitrogen-use efficiency, NUE) for the Himalayan forbs that were examined in situ were similar to the NUE values reported for lowland herbaceous species examined under lowland conditions. In contrast to the common belief, theoretical calculations indicate that the decrease in the rate of photosynthesis due to low atmospheric pressure is small. These Himalayan forbs appeared to overcome this small disadvantage by increasing stomatal conductance. Suppression of photosynthesis caused by blockage of stomata by raindrops appeared to be avoided by either of two mechanisms: plants had large hypostomatous leaves that expanded horizontally or they had obliquely oriented amphistomatous leaves without bundle sheath extensions. All these observations indicate that the gas-exchange characteristics of alpine forbs in the Eastern Himalaya are adapted to the local wet and humid monsoon conditions and thus photosynthetic rates attained during the monsoon period are similar to those of lowland plants.