Effects of chlorantraniliprole on development and reproduction of beet armyworm, <Emphasis Type="Italic">Spodoptera exigua</Emphasis> (Hübner)

The lethal and sublethal effects of chlorantraniliprole against Spodoptera exigua (Hübner) were evaluated under laboratory conditions by oral exposure of neonate larvae to the compound. The 72 h LC₅₀ value of this insecticide to S. exigua was found to be 12.747 μg l⁻¹. A progressive larval mortality of 24.32% for LC₃₀ treatment and 42.61% for LC₅₀ treatment was observed from 4th to 6th day after exposure, which resulted in the reduced pupation rates in exposure groups. The sublethal effects of this chemical were indicated by prolongation of larval period, the increase of pupal weight and decrease in hatch rate of egg. Chlorantraniliprole at LC₃₀ and LC₅₀ rate significantly delayed larval development; the developmental duration of surviving larvae was extended for 22.5 and 28.6%, respectively, compared with that of control group. LC₃₀ treatment increased the mean weight of pupa and induced to the production of heavier pupa (>150 mg). In LC₅₀ treatment, heavier pupa also showed up but the mean weight of pupa was not influenced. The egg hatch rate in LC₅₀ group was significant lower than that in control and LC₃₀ groups. No significant differences in pupal duration, emergence rate, sex ratio, egg number per female, and longevity of adults were observed among treatments. Chlorantraniliprole had exceptional activity against S. exigua according to concentration–response bioassay in laboratory, and the toxicities were primarily resulted from immediate lethality.     

Activities of some defense proteins associated with age or plant family in larch needles of <Emphasis Type="Italic">Larix olgensis</Emphasis> and <Emphasis Type="Italic">L. kaempferi</Emphasis> × <Emphasis Type="Italic">L. gmelinii</Emphasis>

Defence proteins are a kind of chemical defence compounds. They play a key role in plant restraining biotic and abiotic harm. To illuminate activities of some defense proteins associated with age or plant family of larch, the larch needles were collected from two different families of Korean larch, Larix olgensis and a hybrid larch, L. kaempferi × L. gmelinii, respectively, and then the activities of defence proteins in those were tested using a UV spectrophotometry. The results showed that the activities of protective enzymes at the16-/17- and 19-year-age groups were higher than those at the other age groups in the both larch species. While the activities of polyphenol oxidase (PPO) at the 16-/17-year-age group and phenylalanine ammonia-lyase (PAL) and protease inhibitors at the 19-year-age group were the highest among all age groups. Then, compared with L. olgensis, the hybrid larch had significant effects on the activities of protective enzymes. The effects of plant family on the activities of PAL and chymotrypsin inhibitor were significantly different, and then those on the activities of PPO were not significantly different. The effects of the two families in L. olgensis on the activities of trypsin inhibitor (TI) were significantly different, while those in the hybrid larch on the activities of TI were contrary. To conclusions showed that the different age and plant family significantly affected the activities of defence proteins in the needles of two larch species, and then enhanced the larch resistance to pests. These could play a key function in forestry tree genetic improvement and management in future.     

Micropropagation of <Emphasis Type="Italic">Eucalyptus grandis</Emphasis> × <Emphasis Type="Italic">E. urophylla</Emphasis> AEC 224 clone

Genetic transformation systems require protocols that allow regenerating transgenic plants from transformed tissues. This study aimed to establish a protocol for indirect organogenesis in leaf explants of a Eucalyptus grandis  ×  E. urophylla AEC 224 clone. During callogenesis stage, several concentrations of NAA and then NAA or 2,4-D combined with TDZ were tested in JADS culture medium for 30 days, followed by subculture of the explants in the regeneration medium, containing 5.0 µM BA and 0.5 µM NAA for another 30 days. In these media, the explant oxidation rate was high (95 %). Thus, in order to reduce oxidation, different culture media were compared: WPM, MS, JADS and modified QL, followed by explant transfer onto regeneration medium. The highest percentage of regeneration and the lowest oxidation rate were achieved on WPM medium. Then, NAA and 2,4-D were tested in combination with TDZ and also TDZ and BA combined with NAA in WPM medium. The most efficient culture media in terms of shoot regeneration were WPM supplemented with 0.25 µM TDZ and 0.1 µM NAA during 30 days for callus induction and then with 5.0 µM BA and 0.5 µM NAA for another 30 days. This protocol yielded a regeneration rate of 43 %, with a low oxidation of tissues. A rooting experiment was conducted using half strength MS medium and comparing three concentrations of IBA (2.46, 4.90 and 7.35 µM). The highest rooting percentage (35 %) was obtained on medium containing 2.46 µM IBA. Once the shoots were rooted, acclimatization in a greenhouse was not challenging and plant survival reached 100 %.     

Comparative microsporogenesis and flower development in <Emphasis Type="Italic">Eucalyptus urophylla</Emphasis> × <Emphasis Type="Italic">E. grandis</Emphasis>

Microsporogenesis and flower development in Eucalyptus urophylla × E. grandis were examined using chromosome tableting to provide a method to predict the meiotic stages in this species. Although microsporogenesis was normal, cytokinesis during meiosis of pollen mother cells occurred simultaneously, with strong asynchronism observed in the two different lengths of stamens in a flower bud. In a single flower, the developmental period of microsporogenesis in anthers on the longer stamens was always ahead of those on the shorter stamens. Flower development was also asynchronous at different locations on a branch. Flower buds on the upper side of the branch were larger in diameter than those on the lower side. In addition, a correlation was observed between microsporogenesis development and flower bud diameter growth. The pachytene stage was first observed when the diameter of the flower buds increased to 3.0 mm, and the majority of the meiotic stages were observed when bud diameters ranged from 3.5 to 5.0 mm. This study showed that the developmental stages of microsporogenesis in Eucalyptus urophylla × E. grandis could be distinguished readily, which may be applicable to future breeding studies.     

Seasonal variation of selected trace elements in rare endemic species <Emphasis Type="Italic">Thuriferous Juniper</Emphasis> growing in the <Emphasis Type="Italic">Aurès</Emphasis> Mountains of Algeria

We used energy dispersive X-ray fluorescence (EDXRF) to determine the seasonal variation of selected trace elements (Cr, Mn, Zn, Cu, Se and Fe) and some potential toxic elements (Cd, Pb and Br) in Juniperus thurifera subsp. africana Maire (Cupressaceae) a rare medicinal tree, growing indigenously in Aurès Mountains of Algeria. The precision of the results was assessed by analyzing the certified reference material IPE44 (WEPAL) grass leaves. Results showed J. thurifera was characterized by high Cr and Mn contents. During autumn and winter mineral concentrations were higher in general. The levels of Cr, Se and Mn were higher during autumn and winter than during spring and summer. Zn contents were higher during summer than in other seasons. Cu content did not vary by season. The potential toxic elements in J. thurifera (Pb > Cd > Br) were below the permissible limits recommended by the Joint WHO/FAO guidelines except for Pb in autumn and winter.     

A new multistage dynamic model for biological control exemplified by the host–parasitoid system <Emphasis Type="Italic">Spodoptera exigua</Emphasis>–<Emphasis Type="Italic">Chelonus oculator</Emphasis>

Over the last few decades, important advances have been made in understanding of host–parasitoid relations and their applications to biological pest control. Not only has the number of agent species increased, but new manipulation techniques for natural enemies have also been empirically introduced, particularly in greenhouse crops. This makes biocontrol more complex, requiring a new mathematical modeling approach appropriate for the optimization of the release of agents. The present paper aimed at filling this gap by the development of a temperature- and stage-dependent dynamic mathematical model of the host–parasitoid system with an improved functional response. The model is appropriate not only for simulation analysis of the efficiency of biocontrol agents, but also for the application of optimal control methodology for the optimal timing of agent releases, and for the consideration of economic implications. Based on both laboratory and greenhouse trials, the model was validated and fitted to the data of Chelonus oculator (F.) (Hym.: Braconidae) as a biological control agent against the beet armyworm, Spodoptera exigua Hübner (Lep.: Noctuidae). We emphasize that this model can be easily adapted to other interacting species involved in biological or integrated pest control with either parasitoid or predator agents.     

Fitness of heat and desiccation tolerant hybrid strains of <Emphasis Type="Italic">Heterorhabditis</Emphasis> <Emphasis Type="Italic">bacteriophora</Emphasis> (Rhabditidomorpha: Heterorhabditidae)

Entomopathogenic nematodes of the genus Heterorhabditis are a suitable alternative to chemical insecticides because of their high control effect against insects in cryptic environments and their environmental safety. The availability of H. bacteriophora hybrid strains with increased tolerance to environmental extremes could be a driving force for a more widespread use of nematode-based plant protection products. However, increase in heat and desiccation tolerance of hybrid strains could compromise their fitness regarding virulence, host penetration and reproductive capacity. The fitness of heat and desiccation tolerant hybrid strains was compared to the commercial strain EN 01. Only the heat tolerant strains were superior or similar in fitness to strain EN 01. The strains with increased desiccation tolerance were usually less fit, a possible result of a trade-off effect of selection for desiccation tolerance. Hybrid strains selected for enhanced tolerance after an adaptation to stress were generally better in fitness ranking compared to those for which adaptation prior to stress exposure was excluded. This might be a result of pleitropy. Host penetration and virulence was not correlated. The commercial strain had the highest reproduction per mean number of nematodes penetrating the host insect, which is a result of automatic selection of inbred lines with high reproduction potential during the commercial production process in liquid culture.     

Soil microorganisms alleviate the allelopathic effect of <Emphasis Type="Italic">Eucalyptus grandis</Emphasis> × <Emphasis Type="Italic">E</Emphasis>. <Emphasis Type="Italic">urophylla</Emphasis> leachates on <Emphasis Type="Italic">Brassica chinensis</Emphasis>

Soil microbes may be critical players in determining the allelopathic potential of some plants. Low levels of plant community biodiversity in Eucalyptus plantations have been attributed to the allelopathic potential of these tree species. The role of soil microbes in the allelopathic effect of leaf leachates of the hybrid tree Eucalyptus grandis × E. urophylla, was tested in Petri dish assays with Brassica chinensis as a receiver plant. Soils were collected from either a local garden (soil A) or a Eucalyptus plantation (soil B) and half of each soil was sterilized to remove microbes. These soils were then treated with E. grandis × E. urophylla leachates for 0–72 h. Seed germination of B. chinensis was significantly inhibited in soils treated with leaf leachates relative to untreated soils. The inhibitory effect of the leaf leachates was more pronounced in sterilized soils. Total phenolic content was obvious lower in nonsterile leachate-treated soils than in sterile soils. Biomass of B. chinensis was negatively correlated with the total phenolic content in soils. Our findings suggest that soil microbes can alleviate the allelopathic potential of Eucalyptus and thereby its negative impact on plant growth.     

AFLP fingerprinting of <Emphasis Type="Italic">Populus deltoides</Emphasis> and <Emphasis Type="Italic">Populus </Emphasis>×<Emphasis Type="Italic"> canadensis</Emphasis> elite accessions

Amplified fragment length polymorphism (AFLP) was used in genetic fingerprinting of 22 elite Chinese Populus deltoides and Populus × canadensis accessions. The results indicated that each of the nine AFLP primer pairs selected generated fingerprint profiles that were unique to each of the accessions. Therefore, each accession was definitively identified by any of the nine primer pairs. Furthermore, the inter-accession genetic relationships inferred based on 461 polymorphic fragments from the nine AFLP primer pairs were largely consistent with phylogenetic relationships based on morphologic traits. Bootstrap analysis showed that three AFLP primer pairs were required to obtain genetic similarity values with a maximum CV of 10% while 10 AFLP primer pairs could give a maximum CV of 5%. Thus, AFLP can readily be applied for a rapid and accurate evaluation of the degree of similarity between poplar cultivars. In this study, the number of AFLP fragments used was sufficient to establish a reliable estimate of genetic similarity among accessions, with a maximum CV of 5.12%. Therefore, the information on the genetic relationships among the poplar accessions generated in this study in connection with knowledge on agronomic traits may have an impact on poplar breeding and planting in China.     

Quantitative genetics of cold hardiness and growth in <Emphasis Type="Italic">Eucalyptus</Emphasis> as estimated from <Emphasis Type="Italic">E.</Emphasis><Emphasis Type="Italic">urophylla</Emphasis> × <Emphasis Type="Italic">E. tereticornis</Emphasis> hybrids

Interspecific hybrids of Eucalyptus urophylla S. T. Blake × E. tereticornis Smith were used for quantitative genetic analysis of cold hardiness (CH) and growth traits height (H), diameter at breast height (D) and volume (V) at 0.5, 1.5, 2.5 and 4 years of age. The effect of E. tereticornis male variance was significant (P < 0.05) for CH as well as 2.5- and 4-year-old growth except for H ₄, and E. urophylla female × E. tereticornis male interaction was significant for 0.5- and 1.5-year-old growth and CH except for CH _0.5, whereas E. urophylla female effect was not significant for the majority of traits analyzed. Both additive and dominance gene action were present in the genetics of all the traits measured at most ages, indicating that those traits may be improved by hybrid breeding. The σ _D ²/σ _A ² ratios tended to decline after 1.5 years of age in CH and growth traits, implying that breeding value predictions derived from older performance might be more reliable. Estimates of narrow-sense heritability (h ² ) for CH and growth ranged from 0.03 ± 0.05 to 0.28 ± 0.13, depending on trait and age. Additive genetic correlations between CH and V were 0.28 ± 0.48 and 0.69 ± 0.30 at ages 1.5 and 2.5, respectively, suggesting that selection for growth would cause a favorable increase in cold hardiness in E. urophylla × E. tereticornis hybrids.     

Genetic parameters for growth and wood chemical properties in <Emphasis Type="Italic">Eucalyptus urophylla</Emphasis> × <Emphasis Type="Italic">E. tereticornis</Emphasis> hybrids

Key message

Growth and wood chemical properties are important pulpwood traits. Their narrow-sense heritability ranged from 0.03 to 0.49 in Eucalyptus urophylla × E. tereticornis hybrids, indicating low to moderate levels of genetic control. Genetic correlations were mostly favorable for simultaneous improvement on growth and wood traits. Additive and non-additive genetic effects should be considered in making a hybrid breeding strategy.

Context

Eucalypt hybrids are widely planted for pulpwood production purposes. Genetic variations and correlations for growth and wood chemical traits remain to be explored in Eucalyptus interspecific hybrids.

Aims

Our objectives were to clarify the heritability of growth and wood chemical traits and determine the genetic correlations between traits and between trials in E. urophylla × E. tereticornis hybrids.

Methods

Two trials of 59 E. urophylla × E. tereticornis hybrids derived from an incomplete factorial mating design were investigated at age 10 for growth (height and diameter) and wood chemical properties (basic density, cellulose content, hemi-cellulose content, lignin content, and syringyl-to-guaiacyl ratio). Mixed linear models were used to estimate genetic parameters.

Results

Narrow-sense heritability estimates were 0.13?0.22 in growth and 0.03?0.49 in wood traits, indicating low to moderate levels of additive genetic control. Genetic correlations were mostly positively significant for growth with basic density and cellulose content but negatively significant with hemi-cellulose and lignin contents, being favourablefavorable for pulpwood breeding purpose. Type-B correlations between sites were significant for all the traits except diameter and lignin content.

Conclusion

Hybrid superiority warrants the breeding efforts. An appropriate breeding strategy should be able to capture both additive and non-additive genetic effects.

     

Tree and stand growth for clonal <Emphasis Type="Italic">E. urophylla</Emphasis> × <Emphasis Type="Italic">grandis</Emphasis> across a range of initial stockings in southern China

A large initial stocking × fertilizer factorial field trial established in 1993 with a Eucalyptus urophylla × grandis hybrid clone in southern Guangxi province, China, included 6 initial stocking treatments ranging from 667 to 2,222 trees ha⁻¹ and 6 fertilizer treatments. Growth data was collected at various intervals up to age 192 months and also data on outer-wood pilodyn penetration (as an indicator of wood density) and acoustic velocity through the outer-wood (as an indicator of modulus of elasticity) at the last assessment. This report examines the results for the main effect of initial stocking treatments and stocking × fertilizer interactions across ages. There were significant differences between initial stockings for both average individual tree volume and standing volume ha⁻¹ at all ages up to 144 months, but just average individual tree volume at 192 months. At ages up to 75 months total standing volume decreased as initial stocking decreased—maximum volume being obtained at 2,222 trees ha⁻¹. From 88 to 110 months the highest standing volume was obtained at 1,667 trees ha⁻¹ and then at ages 144 and 192 months, 833 trees ha⁻¹ provided the highest standing volume. Average individual tree volume at all ages generally increased as initial stocking decreased—the largest trees were obtained at 667 trees ha⁻¹. As age increased the proportional differences between tree volumes at higher and lower stockings increased—at age 27 months average individual tree volume at 667 trees ha⁻¹ was 72% greater than that at 2,222 trees ha⁻¹ and differences increased steadily with age up to 190% at 192 months. There were no significant differences between initial stockings for the properties of the outer-wood assessed by pilodyn penetration and acoustic velocity. Also, no significant interactions were found between initial stocking and fertilizer treatments for any traits at any ages implying that effects of initial stocking and fertilizer are additive for the traits assessed.     

Midgut cysteine-proteinase activity in the velvetbean caterpillar (<Emphasis Type="Italic">Anticarsia gemmatalis</Emphasis> (Hübner))

Proteinase inhibitors are currently targeted as potential insect control agents, but adaptation to proteinase inhibitors is a recognized limitation to such approach requiring the understanding of how phytophagous species can cope with such compounds. The velvetbean caterpillar (Anticarsia gemmatalis) is a key pest of soybean and well-adapted to its host proteinase inhibitors, which is rich in serine-proteinase inhibitors, particularly trypsin-like proteinase inhibitors. As the expression of cysteine proteinases in the midgut of the velvetbean caterpillar is a potential adaptation to circumvent its host defense, we assessed and characterized the digestive cysteine-proteinase activity from velvetbean caterpillars. Significant soluble and membrane-bound proteolytic activity was obtained and was consistent with those of cysteine proteinases based on the substrate and inhibitors used for their characterization. The K _m and V _max values obtained were 2.35 ± 0.50 mM and 40.89 ± 6.68 nmol min⁻¹ mg⁻¹ for the soluble proteinases and 0.33 ± 0.03 mM and 24.54 ± 0.67 nmol min⁻¹ mg⁻¹ for the membrane-bound proteinases, range of values also consistent with cysteine proteinases. Therefore, the proteolytic activity observed in the velvetbean caterpillar midgut is consistent with that of cysteine proteinases providing preliminary support for the contention of their potential involvement mitigating the negative effects of serine-protease inhibitors in this species.     

Differences in leaf functional traits between red and green leaves of two evergreen shrubs <Emphasis Type="Italic">Photinia</Emphasis> × <Emphasis Type="Italic">fraseri</Emphasis> and <Emphasis Type="Italic">Osmanthus fragrans</Emphasis>

Leaf functional traits are adaptations that enable plants to live under different environmental conditions. This study aims to evaluate the differences in leaf functional traits between red and green leaves of two evergreen shrubs Photinia × fraseri and Osmanthus fragrans. Specific areas of red leaves are higher than that of green leaves in both species. Thus, the material investment per unit area and per lamina of red leaves is significantly lower than that of green leaves, implying an utmost effort of red leaves to increase light capture and use efficiency because of their low leaf-chlorophyll concentration. The higher petiole length of green leaves compared with that of red leaves indicates that adult green leaves may have large fractional biomass allocation to support the lamina structures in capturing light with maximum efficiency and obtaining a high growth rate. The high range of the phenotypic plasticity of leaf size, leaf thickness, single-leaf wet and dry weights, and leaf moisture of green leaves may be beneficial in achieving efficient control of water loss and nutrient deprivation. The high range of phenotypic plasticity of leaf chlorophyll concentration of red leaves may be advantageous in increasing resource (especially light) capture and use efficiency because this leaf type is juvenile in the growth stage and has low leaf-chlorophyll concentration.     

Effects of Thidiazuron,basal medium and light quality on adventitious shoot regeneration from <Emphasis Type="Italic">in vitro</Emphasis> cultured stem of <Emphasis Type="Italic">Populus alba</Emphasis>×<Emphasis Type="Italic">P. berolinensis</Emphasis>

The effect of Thidiazuron （TDZ）, basal media and light quality on adventitious shoot regeneration from in vitro cultured stem of Populus albaxP berolinensis were determined to establish a high efficiency shoot regeneration system from stem explants of P. alba~P berolinensis. Stems ofPopulus alba~P berolinensis were collected from cultured shoots in vitro derived from dormancy buds of 3-year-old seedlings. The stem explants were cultured on MS medium containing 0.02-mg·L-1NAA （naphthaleneacetic acid）, and 0.1, 0.3, 0.5 and 1.0 mg·L-1 concentrations of TDZ to determine the effect of TDZ on shoot regeneration. Three basal media, i.e. MS, woody plant medium （WPM） and B5, were used to test their influences of different media on adventitious shoot regeneration. Green, red, blue and yellow plastic films in comparison with florescent light as control were used to observe their effects on shoot regeneration. The results showed that differ- ent concentrations of TDZ had an evident influence on shoot regeneration. Lower concentration of TDZ （0.1 mg·L-1） resulted in more ad- ventitious shoot regeneration and higher concentration of TDZ （〉0.1 mg·L-1） inhibited shoot regeneration. Among different media, MS medium exhibited a high efficiency for shoot regeneration, followed by WPM medium, while B5 medium inhibited shoot regeneration. Normal light and yellow light exhibited better effects on shoot regeneration, compared with other light.     

Theoretical analysis of the interrelationships between self-thinning,biomass density,and plant form in dense populations of hinoki cypress (<Emphasis Type="Italic">Chamaecyparis</Emphasis> <Emphasis Type="Italic">obtusa</Emphasis>) seedlings

Interrelationships between self-thinning, biomass density, and plant form were mathematically modeled in relation to stand development in which self-thinning is either not occurring or is occurring. The relationship between biomass density and mean shoot mass is derived as a simple power function at the stage when self-thinning does not occur. When self-thinning occurs, constant biomass density is attained when the 3/2 power law of self-thinning applies and the allometric coefficient is assumed to be 1/3 in the allometry between mean plant height and aboveground mass. The applicability of this mathematical model and the allometric reformulations of the self-thinning exponent were tested using experimental data for dense populations of Chamaecyparis obtusa seedlings during the first 2 years of growth. On the basis of the results of the present model and experimental data, the dependence on competition of the mean height:diameter ratio, mean stem diameter, and leaf biomass density are discussed. As a result, the mean height:diameter ratio was almost asymptotically constant at the latter growth stage in the second-year seedlings, so that the 3/2 power law of self-thinning was held in the present analysis. However, the value of height:diameter ratio will become smaller in older stands, because tree height is considered to be asymptotic with respect to tree age due to hydraulic and other limits. Therefore, the present modeling implies that one of the reasons why the 3/2 power law from a geometric basis has been recently rejected depends on whether or not the height:diamter ratio is constant in older trees.     

Biological studies on the olive leaf moth, <Emphasis Type="Italic">Palpita unionalis</Emphasis> Hübner (Lepid., Pyralidae), and the olive moth, <Emphasis Type="Italic">Prays oleae</Emphasis> Bernard (Lepid., Yponomeutidae)

Abstract The biology of two important lepidopterous pests, Prays oleae Bernard and Palpita unionalis Hübner, was studied. The target pests were reared on olive plants, Oleae europaea L. (Oleaceae) under laboratory conditions for two successive generations, from March to June 2002. For P. oleae, the duration of larval stage development in the first generation was 21.4 ± 0.18 days at 19.3–20.9 °C, 65–68 % R.H, while in the second generation it was 14.8 ± 0.10 days at 20.8–24.2 °C, 65–69 % R.H. The number of eggs laid per female ranged from 58 to 109 eggs in the first generation, and from 47 to 113 in the second. The larval stage duration of P. unionalis was 16.3 ± 0.12 days at 16.8–22.9 °C, 65–69 % R.H. and 15.5 ± 0.12 days at 21.6–25.5 °C, 66–69 % R.H. for the first and second generations, respectively. Also, the number of eggs laid per female ranged from 630 to 653 eggs, and from 425 to 493 eggs in the second generation under the same previous laboratory conditions. P. oleae laid eggs at night mostly singly on flower-buds, more on the calyx. than on the petals. The larvae mined on leaves and damaged groups of flowers. P. unionalis adults were active at night, laid eggs singly at twilight usually on the lower surface of foliage. Larvae fed on leaves spinning several leaves together to form shelter for the pupa.     

Effect of major inorganic nutrients on <Emphasis Type="Italic">β</Emphasis>-thujaplicin production in a suspension culture of <Emphasis Type="Italic">Cupressus lusitanica</Emphasis> cells

 The optimum conditions for β-thujaplicin production in a Cupressus lusitanica cell suspension culture were investigated. The conditions required for β-thujaplicin production were clearly different from the conditions for cell growth. The initial phosphate concentration and pH did not affect β-thujaplicin production. A total nitrogen source concentration higher than 3.2 mM suppressed production due to the presence of the ammonium ion. β-Thujaplicin production was observed at 95 mg/l without adding the ammonium ion to the medium. Strict control of major inorganic nutrients was not necessary to produce β-thujaplicin. This finding seems to be favorable for future automated production of β-thujaplicin in commercial cell culture plants.     

Effects of water stress on water use efficiency and water balance components of <Emphasis Type="Italic">Hippophae rhamnoides</Emphasis> and <Emphasis Type="Italic">Caragana intermedia</Emphasis> in the soil–plant–atmosphere continuum

Hippophae rhamnoides (Sea buckthorn) and Caragana intermedia (Intermediate peashrub), which are nitrogen-fixing and perennial temperate shrubs, are extensively used in the agroforestry systems in the loess areas of China. In this study, artificially controlled water gradient experiments were carried out with both shrubs, based on four levels of water supply (normal precipitation, slight drought, drought, and extreme drought). The results showed significant impacts of drought on net photosynthesis rate, biomass accumulation, and biomass allocation in both species. Water use efficiency (WUE) varied with different species, scales, and water stress intensities. WUE at the leaf scale (WUEi) was highest under moderate water stress, while the WUE at the community scale (WUEb) decreased with increasing water stress. We observed alteration in the diel and seasonal transpiration of both species in response to water stress. The night-time transpiration accounted for a small but significant proportion of the water balance, and its importance tended to increase with increasing drought. Both species experienced water deficits under all treatments. Soil moisture of H. rhamnoides declined more severely than that of C. intermedia. The ratio of transpiration to evapotranspiration (T/ET) ranged from 50 to 80% for both species in the growing season. The T/ET and WUEi of H. rhamnoides were comparatively smaller under severe water stress but increased under moist conditions. Our data suggest that H. rhamnoides uses water resources more efficiently under favorable water conditions, and C. intermedia shows competitive advantages under drought conditions. Moreover, possible strategies to maintain water balance in the water-limited agroforestry systems are discussed.     

5<Emphasis Type="Italic">Α</Emphasis>-Reductase inhibitory tannin-related compounds isolated from <Emphasis Type="Italic">Shorea laeviforia</Emphasis>

Five tannin-related compounds – gallic acid, flavogallonic acid dilactone, valoneic acid dilactone, gallagyldilactone, ellagic acid – were isolated from the heartwood of Shorea laeviforia, and the inhibitory activity of each against rat liver 5-reductase was evaluated. Valoneic acid dilactone and gallagyldilactone exhibited positive inhibitory activity, but gallic acid and ellagic acid did not. Flavogallonic acid dilactone stimulated 5-reductase activity, even though this compound is structurally similar to valoneic acid dilactone. The kinetic study of valoneic acid dilactone and gallagyldilactone indicated that the inhibitory behavior of 5-reductase were not competitive against the substrate (testosterone) and were partially competitive against the cofactor (NADPH). Additionally, double inhibition analysis of valoneic acid dilactone and NADP⁺ showed synergetic inhibition. These results suggested that neither valoneic acid dilactone nor gallagyldilactone can affect the binding of testosterone but that either compound could interact with an enzyme–NADP⁺ complex to inhibit 5-reductase.