Chlorophyllase (CLH) is a common plant enzyme that catalyzes the hydrolysis of chlorophyll to form chlorophyllide, a more hydrophilic derivative. For more than a century, the biological role of CLH has been controversial, although this enzyme has been often considered to catalyze chlorophyll catabolism during stress-induced chlorophyll breakdown. In this study, we found that the absence of CLH does not affect chlorophyll breakdown in intact leaf tissue in the absence or the presence of methyljasmonate, which is known to enhance stress-induced chlorophyll breakdown. Fractionation of cellular membranes shows that Arabidopsis (Arabidopsis thaliana) CLH is located in the endoplasmic reticulum and the tonoplast of intact plant cells. These results indicate that CLH is not involved in endogenous chlorophyll catabolism. Instead, we found that CLH promotes chlorophyllide formation upon disruption of leaf cells, or when it is artificially mistargeted to the chloroplast. These results indicate that CLH is responsible for chlorophyllide formation after the collapse of cells, which led us to hypothesize that chlorophyllide formation might be a process of defense against chewing herbivores. We found that Arabidopsis leaves with genetically enhanced CLH activity exhibit toxicity when fed to Spodoptera litura larvae, an insect herbivore. In addition, purified chlorophyllide partially suppresses the growth of the larvae. Taken together, these results support the presence of a unique binary defense system against insect herbivores involving chlorophyll and CLH. Potential mechanisms of chlorophyllide action for defense are discussed.

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Carbon autonomy of current-year shoots in flowering, and of current-year shoots plus 1-year-old shoots (1-year-old shoot system) in fruiting of Siberian alder (Alnus hirsuta var. sibirica) was investigated using a stable isotope of carbon, (13)C. The current-year shoot and 1-year-old shoot systems were fed (13)CO(2) and the atom% excess of (13)C in flowers and fruits was determined. The majority of photosynthate allocated to flower buds was originally assimilated in the leaves of the flowering current-year shoots. Of all the current-year shoots on fruiting 1-year-old shoots, only those nearest to the fruits allocated the assimilated photosynthate to fruit maturation. These results indicate that the current-year shoots and 1-year-old shoot systems are carbon-autonomous units for producing flowers and maturing fruits, respectively.

Current shoots, which form the crown of a tree, are specialized in various functions such as crown expansion, reproduction, and assimilation. We examined the temporal and spatial distribution of reproductive shoots in Alnus hirsuta Turcz. var. sibirica (Fischer) C.K. Schn., assessed their direct and indirect costs of reproduction, and explained their distribution in the crown as the reproductive strategy of a current shoot population. The upper and lower limits to the lengths of current shoots for reproductive growth (flower formation) were 40 and 10 cm, respectively. Reproductive 1-year-old shoots produced fewer shoots in the following year than non-reproductive 1-year-old shoots. In current shoots longer than 40 cm, the increment of reproductive output in the following year by abandonment of reproduction surpassed the decrement of reproductive output in the current year by abandonment of reproduction. This may be one reason for the upper limit of reproductive shoot length. Thus, the current shoot population of A. hirsuta var. sibirica may be divided into three functionally specialized subpopulations: reproductive, maintenance, and exploratory. This specialization is considered to be a reproductive strategy to maximize their lifetime reproductive success.Key words: current shoot population, reproductive ecology, functional specialization, cost of reproduction, Japanese alder.

Structural characteristics of Abies mariesii M.T. Mast. saplings growing in sun and shade in a snowy subalpine parkland in central Japan were assessed to infer how saplings acclimate to suppression by larger individuals in a conifer clump and to extremely snowy conditions. Sun and shade saplings produced structurally different current-year shoots, and allocated biomass to needles and stem differently. Compared with sun saplings, shoots of shade saplings had lower needle mass per unit shoot size, which indicates less dense needle packing and more effective use of the limited available light by avoiding mutual shading among needles. Biomass allocation within lateral branches also differed between sun and shade saplings. Compared with sun saplings, needle mass was a smaller proportion of total branch mass in shade saplings although shade saplings retained needles for longer, thereby compensating, in part, for their lower annual production of needles. Thus shade saplings incur a high mechanical cost to support their low-light acclimated, conspicuously flattened crowns in this snowy habitat. Suppressed saplings are an important component of the persistent conifer clumps in snowy subalpine parklands. The observed structural characteristics of A. mariesii saplings, which ensure high shade- and snow-tolerance, contribute to the dominance of the species in snowy subalpine regions in Honshu, Japan.

Current foliage photosynthetic products are largely stored in the below-ground parts for survival and future growth, and about one-third of the resources for seed production may be attained by fruit photosynthesis. Therefore, the trade-off between current seed production and subsequent growth is weak. The cost of seed production may be buffered by sufficient storage in the below-ground organs, effective photosynthesis under high irradiation and self-assimilation ability of fruits.

Takada’s group developed a method for estimating the yearly transition matrix by calculating the mth power roots of a transition matrix with an interval of m years. However, the probability of obtaining a yearly transition matrix with real and positive elements is unknown. In this study, empirical verification based on transition matrices from previous land-use studies and Monte-Carlo simulations were conducted to estimate the probability of obtaining an appropriate yearly transition probability matrix. In 62 transition probability matrices of previous land-use studies, 54 (87%) could provide a positive or small-negative solution. For randomly generated matrices with differing sizes or power roots, the probability of obtaining a positive or small-negative solution is low. However, the probability is relatively large for matrices with large diagonal elements, exceeding 90% in most cases. These results indicate that Takada et al.’s method is a powerful tool for analyzing land-use dynamics.

To estimate nodule biomass of Alnus hirsuta var. sibirica, an N 2 -fixing tree species, we examined the distribution pattern and size structure of nodules in a 17 to 18 year old stand naturally regenerated after disturbance by road construction in Japan. Nodules were harvested within 1 m from the outer edge of stems of plants with different sizes on four occasions from June to October. The diameter of the subtending root at the base of each nodule and nodule dry weight were measured in 20 cm increments outwards from the base of each stem. Horizontal distribution of nodules around each tree varied greatly among tree diameters at 1.3 m (dbh) within the even-aged stand. In particular, smaller individuals had a concentrated distribution of nodules close to the stem. Nodule abundance occurred further from the stems with increasing tree size. Nodule biomass within 1 m from the outer edge of individual stems increased with tree size ([nodule biomass] = 0.442 [dbh] 2.01 , R 2 =0.747, P<0.05). By using the relationship, nodule biomasses were estimated to be 84.1 kg ha −1 . These results suggest that it is necessary to take into account tree size and patterns of tree distribution in nodule biomass estimates.