Effects of exogenous 3-indoleacetic acid and cadmium stress on the physiological and biochemical characteristics of Cinnamomum camphora.

Indoleacetic acid (IAA) is a plant growth regulator that plays an important role in plant growth and development, and participates in the regulation of abiotic stress. To explore the effect of IAA on cadmium toxicity in Cinnamomum camphora, an indoor potted experiment was conducted with one-year-old C. camphora seedlings. The influence of IAA on cadmium accumulation, net photosynthetic rates, respiration, photosynthetic pigments (chlorophyll a, chlorophyll b, total chlorophyll and carotenoids), osmoregulatory substances (proline, soluble sugar and soluble protein) and the malondialdehyde content in C. camphora leaves treated with 30 mg kg-1 cadmium was analysed with or without the addition of 10 mg kg-1 IAA. Cadmium accumulation in the leaves of C. camphora with the addition of exogenous IAA was significantly higher than accumulation during cadmium stress without additional IAA (ca 69.10% after 60 days' incubation). During the culture period, the net photosynthetic rate in C. camphora leaves subjected to cadmium stress without the addition of IAA was up to 24.31% lower than that of control plants. The net photosynthetic rate in C. camphora leaves subjected to cadmium stress and addition of IAA was up to 30.31% higher than that of leaves subjected to cadmium stress without the addition of IAA. Chlorophyll a, total chlorophyll and carotenoid contents in the cadmium-stressed leaves without the addition of IAA were lower than those in the control treatment. The presence of IAA increased the chlorophyll a, total chlorophyll and carotenoid contents relative to the cadmium stress without the addition of IAA. The respiration rate and concentrations of proline, soluble sugar, soluble protein and malondialdehyde in C. camphora leaves subjected to cadmium stress without the addition of IAA were higher than those in the control. The addition of IAA reduced the respiration rate, and the concentrations of proline, soluble sugar, soluble protein and malondialdehyde in C. camphora leaves when compared with the cadmium stress without the addition of IAA. These results indicate that exogenous IAA improves photosynthetic performance and the growth environment of C. camphora by enhancing the net photosynthetic rate, increasing concentrations of osmoregulatory substances, removing reactive oxygen radicals and eliminating potential damage, thereby reducing the toxic effects of cadmium on C. camphora.

Changes of Cinnamomum camphora root characteristics and soil properties under ozone stress in South China.

High O3 exposure affects the forest growth and soil characteristics. Although there is substantial evidence that O3 does impose a stress on forest trees, the effects of O3 on roots and soil of evergreen broad-leaved tree species in South China remain unknown. The effects of ozone (O3) fumigation on the root biomass, root morphology, root nutrient, soil physical, and chemical properties were examined in Cinnamomum camphora seedlings grown under four O3 treatments (charcoal-filtered air (CF) or O3 at 1×, 2× and 4× ambient concentration). O3 significantly decreased root biomass and root carbon (C). Regardless of O3 level, elevated O3 significantly resulted in reduced root surface area, volume, number of forks, and specific root length (SRL). The percentages of fine to total root in terms of root surface area and root volume of seedlings under the CF and 1 × O3 treatments were significantly higher than those of seedlings under the 4 × O3 treatment, indicating that high O3 level impaired the growth performance of fine roots. O3 affected root growth and structures, which increased soil bulk density and reduced soil total porosity and void ratio. The soil pH under all O3 fumigation treatments significantly increased compared with CF treatment, whereas the organic matter significantly decreased. In conclusion, although the increased O3 level enhanced root N and P under 2 and 4 × O3 treatments compared with 1 × O3 treatment as compensation mechanisms to prevent O3-induced decrease in root C gain and root functions, O3 still decreased the root biomass and root tips, and changed the soil physical and chemical properties.

An Efficient Protocol for Plantlet Regeneration via Direct Organogenesis by Using Nodal Segments from Embryo-Cultured Seedlings of Cinnamomum camphora L.

A simple and efficient plantlet regeneration protocol via direct organogenesis was established for camphor tree (Cinnamomum camphora L.). Stem segments with one node (SN explants) from embryo-cultured seedlings (EC seedlings) were used as explants. Murashige and Skoog (MS) medium supplemented with 0.5 mg/L 2, 4-dichlorophenoxyacetic acid and 2.0 mg/L 6-benzyladenine was used to induce cotyledonary embryo germination. This medium was also used for EC seedlings propagation and adventitious bud induction from SN explants. Regenerated plantlets were cultured on hormone-free MS medium for elongation and root induction. The regeneration capability of SN explants was compared by using EC seedling lines established in this research. EC seedling line EL6 exhibited the highest adventitious bud induction frequency (91.7%) and the highest number of buds per responding explant (5.2), which was considered as the most efficient EC seedling line for further gene transformation research.

Non-stomatal limitation to photosynthesis in Cinnamomum camphora seedings exposed to elevated O3.

Ozone (O3) is the most phytotoxic air pollutant for global forests, with decreased photosynthesis widely regarded as one of its most common effects. However, controversy exists concerning the mechanism that underlies the depressing effects of O3 on CO2 assimilation. In the present study, seedlings of Cinnamomum camphora, a subtropical evergreen tree species that has rarely been studied, were exposed to ambient air (AA), ambient air plus 60 [ppb] O3 (AA+60), or ambient air plus 120 [ppb] O3 (AA+120) in open-top chambers (OTCs) for 2 years. Photosynthetic CO2 exchange and chlorophyll a fluorescence were investigated in the second growing season (2010). We aim to determine whether stomatal or non-stomatal limitation is responsible for the photosynthesis reduction and to explore the potential implications for forest ecosystem functions. Results indicate that elevated O3 (E-O3) reduced the net photosynthetic rates (PN) by 6.0-32.2%, with significant differences between AA+60 and AA+120 and across the four measurement campaigns (MCs). The actual photochemical efficiency of photosystem II (PSII) in saturated light (Fv'/Fm') was also significantly decreased by E-O3, as was the effective quantum yield of PSII photochemistry (ΦPSII). Moreover, E-O3 significantly and negatively impacted the maximum rates of carboxylation (Vcmax) and electron transport (Jmax). Although neither the stomatal conductance (gs) nor the intercellular CO2 concentration (Ci) was decreased by E-O3, PN/gs was significantly reduced. Therefore, the observed reduction in PN in the present study should not be attributed to the unavailability of CO2 due to stomatal limitation, but rather to the O3-induced damage to Ribulose-1,5-bisphosphate carboxylase/oxygenase and the photochemical apparatus. This suggests that the down-regulation of stomatal conductance could fail to occur, and the biochemical processes in protoplasts would become more susceptible to injuries under long-term O3 exposure, which may have important consequences for forest carbon and water budget.

Responses of native broadleaved woody species to elevated ozone in subtropical China.

To assess ozone sensitivity of subtropical broadleaved tree species and explore possible underlying mechanisms, six evergreen and two deciduous native species were exposed to either charcoal-filtered air or elevated O(3) (E-O(3), ∼150ppb) for one growing season. Initial visible symptoms in deciduous species appeared much earlier than those in evergreen species. The species which first showed visible symptoms also had the largest reductions in biomass. E-O(3) induced significant decreases in photosynthesis rate, chlorophyll content and antioxidant capacity but a significant increase in malondialdehyde content in two deciduous species and two evergreen species (Cinnamomum camphora and Cyclobalanopsis glauca). Except C. glauca, however, E-O(3) had no significant effects on stomatal conductance (g(s)), total phenols and ascorbate contents. Difference in O(3) sensitivity among all species was strongly attributed to specific leaf mass rather than g(s). It suggests that some subtropical tree species will be threatened by rising O(3) concentrations in the near future.

Impact of elevated O3 on visible foliar symptom, growth and biomass of Cinnamomum camphora seedlings under different nitrogen loads.

The effects of elevated ozone (O(3)) and enhanced nitrogen (N) on the growth and biomass of Cinnamomum camphora, a subtropical evergreen broad-leaved tree species, were investigated. The seedlings, supplied with N (NH(4)NO(3) solution) at 0, 30 and 60 kg ha(-1) year(-1) (simplified as N0, N30, N60, respectively), were exposed to ambient (AA) or elevated O(3) (AA + 60 nmol mol(-1) and AA + 120 nmol mol(-1), designated as AA + 60 and AA + 120, respectively) for the 2009 and 2010 growing seasons. Symptomatic leaf percentages were significantly increased with O(3) concentration. AA + 120 significantly decreased the mean leaf size and chlorophyll content of both 2009- and 2010-emerged leaves, inhibited the growth of stem height and basal diameter, and reduced biomass accumulation of all plant parts except for leaves. By comparison, only the chlorophyll content of 2009-emerged leaves and root dry weight were significantly reduced under AA + 60. Specific leaf area, total leaf area and foliar biomass were not affected even at a higher O(3) level. On the other hand, N loads, especially N60, exerted significantly positive effects on all variables except mean leaf size and shoot/root ratio. No significant interactions between O(3) and N were detected, suggesting that the N supply at ≤60 kg ha(-1) year(-1) did not significantly modify the response of C. camphora to O(3) in terms of seedling growth and biomass accumulation.

Transient proliferation of proanthocyanidin-accumulating cells on the epidermal apex contributes to highly aluminum-resistant root elongation in camphor tree.

Aluminum (Al) is a harmful element that rapidly inhibits the elongation of plant roots in acidic soils. The release of organic anions explains Al resistance in annual crops, but the mechanisms that are responsible for superior Al resistance in some woody plants remain unclear. We examined cell properties at the surface layer of the root apex in the camphor tree (Cinnamomum camphora) to understand its high Al resistance mechanism. Exposure to 500 µm Al for 8 d, more than 20-fold higher concentration and longer duration than what soybean (Glycine max) can tolerate, only reduced root elongation in the camphor tree to 64% of the control despite the slight induction of citrate release. In addition, Al content in the root apices was maintained at low levels. Histochemical profiling revealed that proanthocyanidin (PA)-accumulating cells were present at the adjacent outer layer of epidermis cells at the root apex, having distinctive zones for cell division and the early phase of cell expansion. Then the PA cells were gradually detached off the root, leaving thin debris behind, and the root surface was replaced with the elongating epidermis cells at the 3- to 4-mm region behind the tip. Al did not affect the proliferation of PA cells or epidermis cells, except for the delay in the start of expansion and the accelerated detachment of the former. In soybean roots, the innermost lateral root cap cells were absent in both PA accumulation and active cell division and failed to protect the epidermal cell expansion at 25 µm Al. These results suggest that transient proliferation and detachment of PA cells may facilitate the expansion of epidermis cells away from Al during root elongation in camphor tree.

Effects of 60-day NO2 fumigation on growth, oxidative stress and antioxidative response in Cinnamomum camphora seedlings.

OBJECTIVE: To study the oxidative stress and antioxidative response of Cinnamomum camphora seedlings exposed to nitrogen dioxide (NO(2)) fumigation. METHODS: Measurements were made up of the growth, chlorophyll content, chlorophyll fluorescence, antioxidant system and lipid peroxidation of one-year-old C. camphora seedlings exposed to NO(2) (0.1, 0.5, and 4 microl/L) fumigation in open top chambers over a period of 60 d. RESULTS: After the first 30 d, 0.5 and 4.0 microl/L NO(2) showed insignificant effects on the growth of C. camphora seedlings. However, exposure to 0.5 and 4.0 microl/L NO(2) for 15 d significantly reduced their chlorophyll content (P<0.05), enhanced their malondialdehyde (MDA) content and superoxide dismutase (SOD) activity (P<0.05), and also significantly reduced the maximal quantum yield of PSII in the dark [the ratio of variable fluorescence to maximal fluorescence (F(v)/F(m))] (P<0.05). In the latter 30 d, 0.5 microl/L NO(2) showed a positive effect on the vitality of the seedlings, which was reflected by a recovery in the ratio of F(v)/F(m) and chlorophyll content, and obviously enhanced growth, SOD activity, ascorbate (AsA) content and glutathione reductase (GR) activity (P<0.05); 4.0 microl/L NO(2) then showed a negative effect, indicated by significant reductions in chlorophyll content and the ratio of F(v)/F(m), and inhibited growth (P<0.05). CONCLUSION: The results suggest adaptation of C. camphora seedlings to 60-d exposure to 0.1 and 0.5 microl/L NO(2), but not to 60-d exposure to 4.0 microl/L NO(2). C. camphora seedlings may protect themselves from injury by strengthening their antioxidant system in response to NO(2)-induced oxidative stress.

[Effects of NO2 on Cinnamomum camphora seedlings growth and photosynthesis].

A 2-month fumigation experiment was conducted with opened top chambers to study the effects of different concentration (0.1, 0.5, and 4.0 microl x L(-1)) NO2 on the growth and photosynthesis of 1-year Cinnamomum camphora seedlings. Fumigation with 0.1 and 0.5 microl NO2 x L(-1) promoted the growth of the seedlings, while with 4.0 microl NO2 x L(-1) was in adverse. The diurnal variation of net photosynthetic rate (P(n)) presented two-peaks, with an obvious depression in midday. 0.5 microl NO2 x L(-1) increased the P(n), the maximum of P(n) reached 8.542 micromol CO2 x m(-2) s(-1); 4.0 microl NO2 x L(-1) decreased the P(n) in most period of time; while the effect of 0.1 microl NO2 x L(-1) varied with time period. Fumigation with 0.5 and 4.0 microl NO2 x L(-1) increased the maximal and minimal values of stomatal conductance (G(s)) and intercellular CO2 concentration (C( i)), while 0.1 microl NO2 x L(-1) increased the maximal and minimal values of C(i) but decreased the maximal and minimal values of G(s). At the middle and late stages of fumigation, the mean P(n) of the seedlings treated with 0.5 microl NO2 x L(-1) was significantly higher than that treated with 0.1 and 4 microl NO2 x L(-1). At the early stage of fumigation, 0.5 and 4.0 micro NO2 x L(-1) significantly decreased the maximal PS II efficiency (F(v)/F(m)); and at the late stage, 4.0 microl NO2 x L(-1) still decreased the F(v)/F(m) significantly.