Seasonal pigment fluctuation in diploid and polyploid Arabidopsis revealed by machine learning-based phenotyping method PlantServation.

Long-term field monitoring of leaf pigment content is informative for understanding plant responses to environments distinct from regulated chambers but is impractical by conventional destructive measurements. We developed PlantServation, a method incorporating robust image-acquisition hardware and deep learning-based software that extracts leaf color by detecting plant individuals automatically. As a case study, we applied PlantServation to examine environmental and genotypic effects on the pigment anthocyanin content estimated from leaf color. We processed >4 million images of small individuals of four Arabidopsis species in the field, where the plant shape, color, and background vary over months. Past radiation, coldness, and precipitation significantly affected the anthocyanin content. The synthetic allopolyploid A. kamchatica recapitulated the fluctuations of natural polyploids by integrating diploid responses. The data support a long-standing hypothesis stating that allopolyploids can inherit and combine the traits of progenitors. PlantServation facilitates the study of plant responses to complex environments termed "in natura".

Canopy structure and phenology modulate the impacts of solar radiation on C and N dynamics during litter decomposition in a temperate forest.

Decomposition of plant organic matter plays a key role in the terrestrial biogeochemical cycles. Sunlight has recently been identified as an important contributor to carbon [C] turnover through photodegradation, accelerating decomposition even in forest ecosystems where understorey solar irradiance remains relatively low. However, it is uncertain how C and nutrients dynamics respond to fluctuations in solar spectral irradiance caused by canopy structure (understorey vs. gaps) and season (open vs. closed canopy phenology). Spectral-attenuation treatments were used to compare litter decomposition over eight months, covering canopy phenology, in a temperate deciduous forest and an adjacent gap. Exposure to the full spectrum of sunlight increased the loss of litter C and lignin by 75% and 64% in the forest gap, and blue light was responsible for respectively 27% and 42% of that loss. Whereas in the understorey, C and lignin loss were similar among spectral-attenuation treatments over the experimental period, except prior to and during spring canopy flush when exposure to the full spectrum of sunlight promoted C loss by 15% overall, 80% of which was attributable to ultraviolet-B (UV-B) radiation. Nitrogen [N] was immobilized in the understorey during canopy flush before the canopy completely closed but N was swiftly released during canopy leaf-fall. Our study suggests that blue-driven photodegradation plays an important role in lignin decomposition and N dynamics in canopy gaps, whereas seasonal canopy phenology affecting sunlight reaching the forest floor drastically changes patterns of C and N in litter during decomposition. Hence, including sunlight dynamics driven by canopy structure and phenology would improve estimates of biogeochemical cycling in forests responding to changes in climate and land-use.

Genetic variation of the relict maple Acer miyabei: uncovering its history of disjunct occurrence and the role of mountain refugia in shaping genetic diversity.

PREMISE: Relict species provide valuable insights into the origin and formation of extant vegetation. Here, we aimed to elucidate the genetic structure and diversity of a riparian relic, Acer miyabei, in Japan. Once widely distributed, it now occurs in three isolated regions. The most northern regional group is located at low elevation on Hokkaido Island, whereas the southernmost group in central Honshu Island is at high elevation in a mountainous landscape. This contrastive distribution enables us to examine the effects of climate oscillations on genetic diversity in relation to topographic variation. METHODS: We collected 604 individuals of A. miyabei from 43 sites. Their genetic structure and diversity were analyzed using 12 microsatellite markers and cpDNA sequences. RESULTS: According to structure analyses, ∆K was lowest at K = 2; the clustering essentially separated many of the individuals in the most northern regional group from the others. In contrast, the two southern groups were not clearly differentiated from each other, despite their geographic discontinuity. The proportion of private alleles was high in populations from the mountain terrain in the southern group although the number of extant populations is limited. CONCLUSIONS: Genetic clustering of A. miyabei is not perfectly congruent with the current patterns of geographic distribution. We infer that disjunction of the two southern groups occurred more recently than that between these groups and the northern group. The mountainous landscape in the most southern region likely provided multiple refugia and contributed to the retention of distinctive genetic variation.

The contribution of photodegradation to litter decomposition in a temperate forest gap and understorey.

Litter decomposition determines carbon (C) backflow to the atmosphere and ecosystem nutrient cycling. Although sunlight provides the indispensable energy for terrestrial biogeochemical processes, the role of photodegradation in decomposition has been relatively neglected in productive mesic ecosystems. To quantify the effects of this variation, we conducted a factorial experiment in the understorey of a temperate deciduous forest and an adjacent gap, using spectral-attenuation-filter treatments. Exposure to the full spectrum of sunlight increased decay rates by nearly 120% and the effect of blue light contributed 75% of this increase. Scaled-up to the whole forest ecosystem, this translates to 13% loss of leaf-litter C through photodegradation over the year of our study for a scenario of 20% gap. Irrespective of the spectral composition, herbaceous and shrub litter lost mass faster than tree litter, with photodegradation contributing the most to surface litter decomposition in forest canopy gaps. Across species, the initial litter lignin and polyphenolic contents predicted photodegradation by blue light and ultraviolet B (UV-B) radiation, respectively. We concluded that photodegradation, modulated by litter quality, is an important driver of decomposition, not just in arid areas, but also in mesic ecosystems such as temperate deciduous forests following gap opening.

Experimental and Field Data Support Range Expansion in an Allopolyploid Arabidopsis Owing to Parental Legacy of Heavy Metal Hyperaccumulation.

Empirical evidence is limited on whether allopolyploid species combine or merge parental adaptations to broaden habitats. The allopolyploid Arabidopsis kamchatica is a hybrid of the two diploid parents Arabidopsis halleri and Arabidopsis lyrata. A. halleri is a facultative heavy metal hyperaccumulator, and may be found in cadmium (Cd) and zinc (Zn) contaminated environments, as well as non-contaminated environments. A. lyrata is considered non-tolerant to these metals, but can be found in serpentine habitats. Therefore, the parents have adaptation to different environments. Here, we measured heavy metals in soils from native populations of A. kamchatica. We found that soil Zn concentration of nearly half of the sampled 40 sites was higher than the critical toxicity level. Many of the sites were near human construction, suggesting adaptation of A. kamchatica to artificially contaminated soils. Over half of the A. kamchatica populations had >1,000 µg g-1 Zn in leaf tissues. Using hydroponic treatments, most genotypes accumulated >3,000 µg g-1 Zn, with high variability among them, indicating substantial genetic variation in heavy metal accumulation. Genes involved in heavy metal hyperaccumulation showed an expression bias in the A. halleri-derived homeolog in widely distributed plant genotypes. We also found that two populations were found growing on serpentine soils. These data suggest that A. kamchatica can inhabit a range of both natural and artificial soil environments with high levels of ions that either of the parents specializes and that it can accumulate varying amount of heavy metals. Our field and experimental data provide a compelling example of combining genetic toolkits for soil adaptations to expand the habitat of an allopolyploid species.

Four-year musculoskeletal examinations among elementary and junior high school students across a single city / Journal of Rural Medicine

Objective: In 2016, Japan started conducting musculoskeletal examinations that included “limb status” of students as an essential item. Our institution implemented a unified musculoskeletal examination for all public elementary and junior high schools in T-city. In this study, we aimed to report the progress in the past 4 years.Patients and Methods: The Tsukuba Childhood Locomotive-Organ Screening Sheet (T-CLOSS), which is a questionnaire that includes some nationwide-recommended questions, was prepared and distributed to students. Results of the questionnaires were analyzed, and the orthopedic surgeon conducted examinations for the extracted items. From these questionnaires, the ratio of each item, rate of advisory for screening, and content of the advisory were investigated.Results: During 2016–2019, musculoskeletal examinations were conducted in every public schools, with nearly 20,000 students in T-city. The consultation advisory rate was 6.7% in 2019. Of the 524 students who received the third screening recommendation, the actual consultation rate was 248 (40.8%). After the third screening, the proportion of students requiring treatment and outpatient visits was 46.7% (n=248), which accounted for 1.2% of all elementary and junior high school students in the city.Conclusion: We reported the results of 4-year musculoskeletal examinations in a city. In our screening, we distributed a uniform questionnaire throughout the city, and orthopedic surgeons performed secondary examinations of identified students in all schools. This appears to be an advanced effort to prevent musculoskeletal impairment in students. We hope to conduct more sophisticated musculoskeletal examinations using our results, aiming at early detection, early treatment, and improvement of musculoskeletal function in elementary and junior high school students.

Determining the relationship between physical status and musculoskeletal injuries in children: a cohort study / Journal of Rural Medicine

Background: In Japan, in 2016, the School Health and Safety Act was revised and examination of extremities in addition to scoliosis became mandatory. Musculoskeletal examinations were subsequently started using a mark sheet-type questionnaire. In the present study, we aimed to analyze the relationship between physical findings and musculoskeletal problems and propose a preventive strategy for musculoskeletal injuries.Methods: In 2017, a total of 4,073 elementary and middle school students underwent direct musculoskeletal examination. In a direct examination, the following elements were included: torticollis; scoliosis; stiffness of the shoulder, elbow, hip, knee, and ankle; flexion and extension in standing position; flat foot; hallux valgus; and alignment of the upper and lower extremities. Of the 4,073 students who underwent direct examination in early 2017, only 3,754 were able to complete the mark sheet-type questionnaires in early 2018. A prospective longitudinal analysis of the data gathered was performed.Results: A total of 396 (11%) students had injuries. The ankle sprain/non-ankle sprain group comprised 119 (3%)/3,635 (97%) students, while the fracture/non-fracture group comprised 105 (2.8%)/3,650 (97.2%) students, respectively. Comparing the sprain group with the non-sprain group, ankle stiffness significantly correlated with ankle sprain in the univariable and multivariable analyses. Injuries occurred more frequently among boys, older students, students with stiff bodies, and students who were involved in sports activities of longer duration.Conclusion: Ankle stiffness was assumed to be a risk factor for ankle sprain. Stretching of the ankle might be effective for preventing ankle sprain. However, further interventional studies are needed to confirm this finding.

Factors Influencing Leaf- and Root-Associated Communities of Bacteria and Fungi Across 33 Plant Orders in a Grassland.

In terrestrial ecosystems, plants interact with diverse taxonomic groups of bacteria and fungi in the phyllosphere and rhizosphere. Although recent studies based on high-throughput DNA sequencing have drastically increased our understanding of plant-associated microbiomes, we still have limited knowledge of how plant species in a species-rich community differ in their leaf and root microbiome compositions. In a cool-temperate semi-natural grassland in Japan, we compared leaf- and root-associated microbiomes across 137 plant species belonging to 33 plant orders. Based on the whole-microbiome inventory data, we analyzed how sampling season as well as the taxonomy, nativeness (native or alien), lifeform (herbaceous or woody), and mycorrhizal type of host plants could contribute to variation in microbiome compositions among co-occurring plant species. The data also allowed us to explore prokaryote and fungal lineages showing preferences for specific host characteristics. The list of microbial taxa showing significant host preferences involved those potentially having some impacts on survival, growth, or environmental resistance of host plants. Overall, this study provides a platform for understanding how plant and microbial communities are linked with each other at the ecosystem level.

Prophylactic pinning should be considered in patients at risk for slipped capital femoral epiphysis / Journal of Rural Medicine

Background: The use of prophylactic contralateral pinning for slipped capital femoral epiphysis (SCFE) remains controversial. This study evaluated the outcome of SCFE treatment and examined the use of prophylactic pinning.Methods: The study included 44 patients (33 men, 11 women; 54 hips [right, 31; left, 23]), with mean age of 12.9 (7.3–29) years, who underwent treatment between 1986 and 2017, with follow-up for more than 6 months. Patients were divided into 3 groups: group 1 had bilateral SCFE at first presentation, group 2 developed contralateral side SCFE during follow-up, and group 3 had unilateral SCFE until final follow-up. Three patients who received prophylactic pinning were excluded. Univariate and multivariate logistic analyses were performed.Results: Overall, 93% (50/54) of hips underwent positional reduction and in situ fixation and 7.4% (4/54) underwent open reduction. Mean follow-up period was 4.8 (0.5–25) years. Groups 1, 2, and 3 had 7, 3, and 31 cases, respectively. Sex, age, and follow-up period showed no significant differences among the groups. The Rohrer index was significantly higher in group 1, the affected side posterior sloping angle (PSA) was significantly higher in group 3, and the contralateral side PSA and percentage with endocrinopathy were significantly higher in group 2. In multivariate logistic analysis, age, sex, Rohrer index, affected side PSA, and endocrinopathy were significantly correlated with bilateral SCFE.Conclusion: We recommend prophylactic contralateral side pinning in patients with risk factors of obesity, high PSA before slipping, and endocrinopathy. Careful observation until growth plate closure is required in patients without risk factors.

Relationship between exercise time and musculoskeletal problems in children / Journal of Rural Medicine

Objectives: The purpose of this study was to evaluate the relationship between exercise time and musculoskeletal problems and to determine the appropriate amount of exercise for children in both lower- and higher-grade levels of elementary and junior high schools.Materials and Methods: Mark-sheet-type questionnaires were distributed to and collected from all elementary and junior high schools in two cities. We collected 22,494 questionnaires in total. The relationship between exercise time and musculoskeletal problems was analyzed. The χ2 test and multivariate logistic regression analysis were used for statistical analyses.Results: The mean exercise time in school, in addition to physical education time, was 3.1 hours per week. In 56% of the children, the exercise time was less than 2 hours per week, and in 13% of the children, the exercise time was more than 10 hours per week. Although the rate of sports injury increased with an increase in exercise time, the duration of one-leg stand (a test of balance and muscle strength) also increased with an increase in exercise time. The cut-off values for sports injuries in boys/girls were 2.9/2.9 hours, 4.0/2.9 hours, and 7.5/4.2 hours in lower grade elementary school, higher grade elementary school, and junior high school, respectively.Conclusions: Although an appropriate amount of exercise improves one’s physical health and ability, excessive exercise leads to musculoskeletal problems. Approximately 7 hours/week of exercise is recommended for junior high school students. In elementary school, the exercise time should be carefully decided as the musculoskeletal system of the students is still immature.

Tissue Culture as a Source of Replicates in Nonmodel Plants: Variation in Cold Response in Arabidopsis lyrata ssp. petraea.

While genotype-environment interaction is increasingly receiving attention by ecologists and evolutionary biologists, such studies need genetically homogeneous replicates-a challenging hurdle in outcrossing plants. This could be potentially overcome by using tissue culture techniques. However, plants regenerated from tissue culture may show aberrant phenotypes and "somaclonal" variation. Here, we examined somaclonal variation due to tissue culturing using the response to cold treatment of photosynthetic efficiency (chlorophyll fluorescence measurements for Fv/Fm, Fv'/Fm', and ΦPSII, representing maximum efficiency of photosynthesis for dark- and light-adapted leaves, and the actual electron transport operating efficiency, respectively, which are reliable indicators of photoinhibition and damage to the photosynthetic electron transport system). We compared this to variation among half-sibling seedlings from three different families of Arabidopsis lyrata ssp. petraea Somaclonal variation was limited, and we could detect within-family variation in change in chlorophyll fluorescence due to cold shock successfully with the help of tissue-culture derived replicates. Icelandic and Norwegian families exhibited higher chlorophyll fluorescence, suggesting higher performance after cold shock, than a Swedish family. Although the main effect of tissue culture on Fv/Fm, Fv'/Fm', and ΦPSII was small, there were significant interactions between tissue culture and family, suggesting that the effect of tissue culture is genotype-specific. Tissue-cultured plantlets were less affected by cold treatment than seedlings, but to a different extent in each family. These interactive effects, however, were comparable to, or much smaller than the single effect of family. These results suggest that tissue culture is a useful method for obtaining genetically homogenous replicates for studying genotype-environment interaction related to adaptively-relevant phenotypes, such as cold response, in nonmodel outcrossing plants.

Conserved but Attenuated Parental Gene Expression in Allopolyploids: Constitutive Zinc Hyperaccumulation in the Allotetraploid Arabidopsis kamchatica.

Allopolyploidization combines parental genomes and often confers broader species distribution. However, little is known about parentally transmitted gene expression underlying quantitative traits following allopolyploidization because of the complexity of polyploid genomes. The allopolyploid species Arabidopsis kamchatica is a natural hybrid of the zinc hyperaccumulator Arabidopsis halleri and of the nonaccumulator Arabidopsis lyrata We found that A. kamchatica retained the ability to hyperaccumulate zinc from A. halleri and grows in soils with both low and high metal content. Hyperaccumulation of zinc by A. kamchatica was reduced to about half of A. halleri, but is 10-fold greater than A. lyrata Homeologs derived from A. halleri had significantly higher levels of expression of genes such as HEAVY METAL ATPASE4 (HMA4), METAL TRANSPORTER PROTEIN1 and other metal ion transporters than those derived from A. lyrata, which suggests cis-regulatory differences. A. kamchatica has on average about half the expression of these genes compared with A. halleri due to fixed heterozygosity inherent in allopolyploids. Zinc treatment significantly changed the ratios of expression of 1% of homeologous pairs, including genes putatively involved in metal homeostasis. Resequencing data showed a significant reduction in genetic diversity over a large genomic region (290 kb) surrounding the HMA4 locus derived from the A. halleri parent compared with the syntenic A. lyrata-derived region, which suggests different evolutionary histories. We also estimated that three A. halleri-derived HMA4 copies are present in A. kamchatica Our findings support a transcriptomic model in which environment-related transcriptional patterns of both parents are conserved but attenuated in the allopolyploids.

The use of a novel in-bed active Leg Exercise Apparatus (LEX) for increasing venous blood flow / Journal of Rural Medicine

<b>Objective:</b> The incidence of pulmonary embolism (PE) and leg deep vein thrombosis (DVT) has increased in recent years in association with aging and an increase in the number of bedridden individuals. We developed an active in-bed leg exercise apparatus labeled the Leg Exercise Apparatus (LEX) for DVT prevention. We compared the effect of leg exercises performed using the LEX to conventional active ankle exercises on increased blood flow.<br><b>Materials & Methods:</b> The subjects were eight healthy adult volunteers [five men and three women, aged 20–34 (mean 27.0) years]. Subjects performed two types of exercise; exercise 1 consisted of leg exercises using the LEX, while exercise 2 consisted of in-bed active plantar flexion/dorsiflexion exercises without the device. Measurements were taken 1, 5, 10, 20, and 30 minutes after exercise including common femoral vein blood flow, mean blood flow velocity, maximum blood flow velocity, and vessel diameter using Doppler ultrasound. Statistical procedures included timed measurement data analysis using a linear mixed model. A Bonferroni correction was used for multiple comparisons.<br><b>Results:</b> Compared to resting levels, blood flow reached a maximum value 1 minute after exercise for both exercise types, with a significantly greater increase after exercise 1 (1.76-fold increase) compared to exercise 2 (1.44-fold increase) (<i>p</i> = 0.005). There was a significant difference (<i>p</i> = 0.03) between the two exercises for all values from 1 minute to 30 minutes following exercise. There was no significant difference between exercises for peak or mean blood flow velocity. Compared to resting levels, blood vessel diameter reached a maximum value of 1.47-fold greater at 5 minutes post-exercise for exercise 1 and a maximum value of 1.21-fold greater at 1 minute post-exercise for exercise 2.<br><b>Conclusions:</b> Exercise using the LEX increased lower leg venous blood flow and vessel diameter. We propose that the LEX may serve as a new DVT prevention tool.

Development and evaluation of microsatellite markers for Acer miyabei (Sapindaceae), a threatened maple species in East Asia.

PREMISE OF THE STUDY: Twelve microsatellite markers were developed and characterized in a threatened maple species, Acer miyabei (Sapindaceae), for use in population genetic analyses. METHODS AND RESULTS: Using Ion Personal Genome Machine (PGM) sequencing, we developed microsatellite markers with perfect di- and trinucleotide repeats. These markers were tested on a total of 44 individuals from two natural populations of A. miyabei subsp. miyabei f. miyabei in Hokkaido Island, Japan. The number of alleles per locus ranged from two to eight. The observed and expected heterozygosities per locus ranged from 0.05 to 0.75 and from 0.05 to 0.79, respectively. Some of the markers were successfully transferred to the closely related species A. campestre, A. platanoides, and A. pictum. CONCLUSIONS: The developed markers will be useful in characterizing the genetic structure and diversity of A. miyabei and will help to understand its spatial genetic variation, levels of inbreeding, and patterns of gene flow, thereby providing a basis for conservation.

Mass flowering of the tropical tree Shorea beccariana was preceded by expression changes in flowering and drought-responsive genes.

Community-level mass flowering, known as general flowering, which occurs in South-East Asia at supra-annual irregular intervals, is considered a particularly spectacular phenomenon in tropical ecology. Recent studies have proposed several proximate factors inducing general flowering, such as drought and falls in minimum temperature. However, limited empirical data on the developmental and physiological processes have been available to test the significance of such factors. To overcome this limitation and test the hypotheses that general flowering is triggered by the proposed factors, we conducted an 'ecological transcriptome' study of a mass flowering species, Shorea beccariana, comparing meteorological data with genome-wide expression patterns obtained using next-generation sequencing. Among the 98 flowering-related genes identified, the homologs of a floral pathway integrator, SbFT, and a floral repressor, SbSVP, showed dramatic transcriptional changes before flowering, and their flowering functions were confirmed using transgenic Arabidopsis thaliana. Expression in drought-responsive and sucrose-induced genes also changed before flowering. All these expression changes occurred when the flowering-inducing level of drought was reached, as estimated using data from the preceding 10 years. These genome-wide expression data support the hypothesis that drought is a trigger for general flowering.

Practical Training at a Hospital Pharmacy Improved the Drug Information Literacy of Pharmacy Students / 医薬品情報学

<b>Objective: </b>Drug information literacy is necessary for pharmacists.  This study investigated the effect of practical training on the drug information literacy of pharmacy students.<br><b>Methods: </b>We conducted practical training in the drug information room of hospital pharmacy.  Then we asked the students two questions before and after the training.  We scored the drug information responses of students using four-point rating scales based on objective criteria.  We compared the scores obtained before and after training.  In addition, we researched the realization of several secondary and tertiary sources using a visual analog scale (VAS).  We then compared VAS findings before and after the training.<br><b>Results: </b>The score for drug information responses was significantly improved.  For sources other than MEDLINE (Pub Med), the VAS score also improved.  The students were aware of the information on package inserts before training.  There was significant improvement in awareness of books about pregnancy and lactation (e.g. Drugs in Pregnancy and Lactation) after training.  However, there were significant individual differences in awareness of MEDLINE, Up To Date, and the Cochrane Library.<br><b>Conclusion: </b>This practical training experience improved the drug information literacy of pharmacy students.  Before training, there were significant deficiencies in the process of literature search.  Therefore, this training was effective for advancement in drug information literacy of pharmacy students.

Variation at range margins across multiple spatial scales: environmental temperature, population genetics and metabolomic phenotype.

Range margins are spatially complex, with environmental, genetic and phenotypic variations occurring across a range of spatial scales. We examine variation in temperature, genes and metabolomic profiles within and between populations of the subalpine perennial plant Arabidopsis lyrata ssp. petraea from across its northwest European range. Our surveys cover a gradient of fragmentation from largely continuous populations in Iceland, through more fragmented Scandinavian populations, to increasingly widely scattered populations at the range margin in Scotland, Wales and Ireland. Temperature regimes vary substantially within some populations, but within-population variation represents a larger fraction of genetic and especially metabolomic variances. Both physical distance and temperature differences between sites are found to be associated with genetic profiles, but not metabolomic profiles, and no relationship was found between genetic and metabolomic population structures in any region. Genetic similarity between plants within populations is the highest in the fragmented populations at the range margin, but differentiation across space is the highest there as well, suggesting that regional patterns of genetic diversity may be scale dependent.

Reproductive success and distance to conspecific adults in the sparsely distributed tree Kalopanax pictus.

To understand how a sparsely distributed species can maintain viable populations, and with a particular interest in distance-dependent reproductive success, we investigated the ovule survival and the factors that determine the survival for sparsely distributed Kalopanax pictus (Araliaceae) in a temperate forest landscape around the Ogawa Forest Reserve, central Japan. We found 154 potentially mature trees (>20 cm diameter at breast height) in a 600-ha site. The year 2000 was a mass flowering year for K. pictus; 96.8% of the trees examined flowered in that year; however, this was less than half of the trees that flowered in other years. The flowers of K. pictus include protandrous inflorescences, which bloom on individual tress in midsummer when few other tree species bloom. We conducted pollination experiments with out-crossed, self-pollinated, bagged, and control inflorescences. Results suggest that self-pollination is limited, partially by low self-compatibility and partially by protandry. Ovule survival from the flower to the seed stages was not dependent on the temporal flowering density of conspecific adults. The flowering habit and presence of effective pollinators may allow K. pictus to avoid the negative effects of sparse distribution on pollination efficiency. However, ovule survival during the ovary development stage depended on infection by a species-specific fungus, Mycosphaerella acanthopanacis. The fungus damages leaves, and the infection intensity depends on the distance between conspecific K. pictus adults. Thus, an advantage of sparse distribution in K. pictus is disease avoidance, whereas its disadvantages are overcome by pollination efficiency.

Multiple factors contribute to outcrossing in a tropical emergent Dipterocarpus tempehes, including a new pollen-tube guidance mechanism for self-incompatibility.

The self-rejection system of Dipterocarpus tempehes (Dipterocarpaceae), an emergent tree of the lowland tropical forests of Borneo, were studied by means of pollination experiments, fluorescence microscopy of pollen tubes, and monitoring of ovary maturation patterns. Fruit set was higher in cross-pollinated flowers than in control and self-pollinated flowers, indicating the existence of pollen limitation and a self-rejection system. Although the adhesion and the germination of self-pollen and the growth of self-pollen tubes were not inhibited, the proportion of cross-pollen tubes that entered the style was 1.7-2.3 times higher than that of self-pollen tubes, indicating a partial self-incompatibility that inhibits self-pollen tubes from entering the style hollow. These results suggest, for the first time, that self-incompatibility is caused by a defect of pollen-tube guidance. We also suggest a threshold effect in number of pollen tubes or late-acting self-incompatibility to fully explain the drastic and uniform selection against self-pollinated flowers before ovary swelling. After that, maternal selection and/or inbreeding depression caused the abortion and delayed maturation of self-pollinated flowers. The advantages of the self-rejection process composed of partial early-acting self-incompatibility and relatively strong delayed abortion is discussed with respect to the general-flowering phenomenon in lowland dipterocarp forests.