Characterization,expression and phylogenetic study of R2R3-MYB genes in orchid

cDNA fragments representing 21 R2R3-MYB genes were isolated by RT-PCR from the Dendrobiumorchid hybrid Woo Leng. Six full-length cDNA clones were obtained from a flower cDNA library, four of which, DwMYB1, DwMYB2, DwMYB8 and DwMYB10, represent typical plant R2R3-MYB genes. The conceptual DwMYB4 protein is truncated at the C-terminal region and contains the R2 repeat and the N-terminal half of the R3 repeat (R2R3). DwMYB4 expression is restricted to flowers. DwMYB9 contains an 8 amino acid N-terminal deletion in the R2 repeat (R2R3) and is expressed at high levels in mature flower and inflorescence, but at very low levels in young flower buds. DwMYB8 and DwMYB10 show similar expression patterns and share very high sequence similarity in the N-terminal part of the MYB domain. Analysis of amino acid substitution indicated that the pattern and type of substitution between Arabidopsis and maize are quite different. Maize may have more conserved substitution in the MYB^BRH domain than Arabidopsis.     

Transfer of pollinaria on birds’ feet: a new pollination system in orchids

We report the discovery of a new mechanism of pollination in orchids: transfer of pollinaria on the feet of birds. Observations carried out in South Africa and Malawi showed that the orchids Disa chrysostachya Sw. and Disa satyriopsis Kraenzl. are pollinated by sunbirds. Pollinaria of these orchids become attached firmly to the birds toes when they perch on the tall narrow inflorescences which are packed tightly with numerous small orange flowers. Birds typically perch on the lower part of an inflorescence while reaching up to feed on nectar in flowers on the upper part, but occasionally reverse this position to probe the lower flowers. The nectar is contained within a short bulbous spur with a narrow entrance that permits entrance of a sunbirds slender tongue. Contrary to expectation, the pollination mechanism in D. chrysostachya is remarkably efficient with about 6.1% of pollen reaching stigmas on other plants and fruit set occurring in 95% of flowers at one site. Birds seldom move their feet once perched, thus minimizing the incidence of self-pollination, either within or between flowers on an inflorescence.     

Importance of woody debris in seed germination of Tipularia discolor (Orchidaceae)

Concerns about declining populations of terrestrial orchids make it important to identify the environmental factors crucial to seedling recruitment. This study shows that seedlings of Tipularia discolor (cranefly orchid) primarily occur on decomposing wood. Extensive searches of decomposing logs and stumps in mature and successional forests revealed seedlings at 24 sites, of which 15 could be identified as originating from seven different deciduous trees and one conifer. Seeds were planted in natural habitats to test the hypothesis that germination requires decomposing wood. In one experiment, seeds were placed into soil at sites where adult plants were abundant; no germination resulted. In a second experiment, germination of seeds sown in ambient soil was compared with sowings in plots amended with decomposing wood collected from a stump where spontaneous seedlings grew. Germination was much more frequent in plots amended with decomposing wood. We conclude that germination of T. discolor is stimulated in substrates that contain decomposing wood; judging from the occurrence of spontaneous seedlings, wood from at variety of tree species offer a suitable substrate.     

A Nectar Reward: Is More Better?1

Characters involved in pollinator attraction are likely maintained by selection. Plants that invest more in floral displays and/or rewards are expected to attract more pollinators than those that do not. A large number of plants, however, are severely pollen-limited yet either produce small rewards or none at all. The orchid, Comparettia falcata, is a pollinator-dependent, self-compatible epiphyte distributed throughout the Greater Antilles, Central and South America. In Puerto Rico where it is pollinated by the hummingbird Chlorostilbon maugaeus, C. falcata presents a smaller nectar reward than most other plants pollinated by the same species. To determine whether or not selection would favor the production of higher nectar levels, we enhanced the quantity of nectar offered by flowers in a Puerto Rican population for two flowering seasons. We monitored visitation frequencies, pollen movement, and reproductive success at three sites with different canopy coverages. Daily censuses of hummingbirds provided estimates of relative pollinator abundance. A multiway contingency test employing Wal?s statistic showed no overall differences in reproductive success between plants with enhanced rewards and unmanipulated controls. Site differences, however, were clear. Plants of the mid- and high-light sites had greater success than those of the low-light site, and the differences were usually at least two-fold. There was a significant site-treatment interaction in reproductive success that could be attributed to the overall trend whereby controls of both mid- and high-light sites did better than the nectar-enhanced plants. Most of the observed pollinations (85%) with stained pollinia resulted in self-pollinations that did not differ among treatments. Seed crops from self- and cross-pollinations revealed no differences in the number of viable seeds. Because we found little evidence of selection for increasing nectar reward via inbreeding depression or male and female reproductive success, and previous studies have indicated that meager natural levels of reward are better than none at all, we suspect that reward production in C. falcata may be driven by a combination of pollination-limitation and resource constraints.     

Effects of irradiation on the reproductive ability of Zonitoides arboreus, a snail pest of orchid roots

The effect of irradiation on the reproductive ability of the orchid snail, Zonitoides arboreus, a serious pest of potted orchids in Hawaii, was investigated. Weekly egg production averaged between 0.8 and 1.9 over a 9–wk period for snails not exposed to irradiation, and egg hatch averaged 61%. In comparison to untreated controls, irradiation of snails at the lowest dose tested (34–37 Gy) reduced egg production and egg hatch by 63% and 94%, respectively over a 9–wk period. None of the snails treated with levels of irradiation ≥ 69 Gy produced viable eggs. This is the first study measuring the effect of ionising irradiation on a terrestrial snail or slug species using sterilising doses. Overall, the results show that the reproductive ability of this snail species is affected by irradiation in a similar manner as for Biomphalaria glabrata, an aquatic snail for which the effects of irradiation have been studied in detail.     

Pollinator limitaton in a deceptive orchid, Pogonia japonica, on a floating peat mat

Deceptive orchid species that offer no floral rewards to pollinators often experience pollinator limitation because they. The breeding system and fruit set were investigated in order to examine pollinator limitation for a population of a deceptive orchid, Pogonia japonica , on a floating peat mat in Mizorogaike Pond, western Japan. Fruit sets for outcross-pollinated, self-pollinated and pollinator-excluded flowers were 75%, 80%, and 0%, respectively. Thus, this species was self-compatible but neither autogamous nor apogamous. The fruit set for open-pollinated flowers was 22.9% and 17.4% in 1994 and 1995, suggesting that pollinator limitation did occur in the field. Larger flowers were more likely to develop into fruit than smaller ones in open pollination (36% vs 8%). Thus, flower size seems to affect fruiting of this species. No effective visits by major pollinators such as bees, which could carry pollinia, were observed during 16.5 h of observation during daytime. Instead, thrips were often found moving on gynostemium or pollinia and circumstantial evidence that suggests thrips may partly transfer pollen in pieces out of a granular pollinium of P. japonica was obtained. Pogonia japonica should come to use thrips as a pollinator to supplement low fruit set under limited flower visitation by major pollinators.     

POLLINATORS DISCRIMINATE AMONG FLORAL HEIGHTS OF A SEXUALLY DECEPTIVE ORCHID: IMPLICATIONS FOR SELECTION

Pollinators have influenced the evolution of many morphological floral traits, although few studies have shown that pollinators have influenced plant height. Chiloglottis trilabra is one of many Australian orchids that deceive and attract male pollinators by mimicking the sex pheromones and morphology of females insects. Orchids in this genus have unusually short flowers whose peduncle elongates dramatically after pollination to approximately twice the original height. In a series of choice experiments in the field, we show that pollinators of C. trilabra strongly discriminate among floral heights, preferring flowers presented at 15 cm-20 cm over flowers presented at lower and higher positions (ranging from 2 cm-100 cm). Our results suggested pollinators have the potential to mediate stabilizing selection for floral height when pollination is limiting. However, the natural height range of the orchid (mean = 10 cm, range 5 cm–15 cm) was lower than the experimentally determined optimum for visitation frequency. This difference may indicate that pollinator-mediated selection does not occur in this species, perhaps because seed set is not sufficiently limited. Alternatively, other life-history factors may counteract pollinator-mediated selection, yielding an evolutionary compromise in height.     

THE GENETIC CONSEQUENCES OF WORKER ANT POLLINATION IN A SELF-COMPATIBLE,CLONAL ORCHID

The self-compatible orchid Microtis parviflora is pollinated by the flightless worker caste of the ant Iridomyrmex gracilis. The orchid is clonal and forms small patches, usually less than 1 m², of disconnected individual ramets. Ant pollinators visited and revisited a limited proportion of available inflorescences, and 40% of all flower visits occurred within plants promoting self-pollination. Pollen labels indicated that self-pollination accounted for 51% of the pollen transfers, although pollen carryover extended beyond 16 flowers on 2 or 3 inflorescences. The distribution of ant movements between plants was leptokurtic with a mean of 12.4 ± 14.9 cm and a maximum of 89 cm, but a high proportion of movements were within clones accentuating the level of self-pollination. However, some pollen transfers between inflorescences of unlike genotypes contributed to a low incidence (max = 8%) of outcrossing. In 12 patches examined by electrophoresis, the density varied from 11 to 61 inflorescences per m² and a maximum of only 4 genotypes were detected. Electrophoretic analysis revealed populations were highly inbred: only 23% (N = 17) of the loci were polymorphic and the mean gene diversity h, was 2.7%. Heterozygotes were observed in only one population given a mean fixation index F, of 0.982. These results reflect the combined effects of restricted ant foraging and clonality. Nevertheless, while ant foraging was restricted, some outcrossing occurred and in the absence of clonality it is likely that ant foraging would have yielded a mixed mating system similar to those reported for a wide array of insect pollinators. Given the ability of ants to generate pollen flow, the reasons for the rarity of ant pollination appear to lie elsewhere.     

TIBA affects the induction of direct somatic embryogenesis from leaf explants of Oncidium

To study the effect of auxin on direct somatic embryogenesis from leaf cultures ofOncidium `Gower Ramsey', 1-cm-long explants have been cultured in vitro testing IAA, 2,4-, quercetin, TIBA and PCIB. On a modified MS medium devoid of plant growth regulators, leaf cells of three regions (leaf tips, adaxial sides and cut ends) formed somatic embryos. After 8 weeks in culture, the frequencies of embryo-forming explants were 55, 52.5 and 30 % on leaf tips, adaxial sides and cut ends, respectively, and the numbers of embryos per dish was 89.3. Except for TIBA, other growth regulators (IAA, 2,4-, quercetin, PCIB) and their combinations tested, all retarded direct embryo formation. In the presence of 0.1 and 0.5 M TIBA, leaf tip, adaxial sides and cuts end of explants gave almost the same embryogenic response as the control. However, 10 and 27.5 % of explants were induced to form embryos from abaxial sides, and these explants did not form embryos on cut ends. In addition, after 8weeks in culture, TIBA at 0.5M highly promoted the mean numbers of embryos per dish to 134.2.