A feasible method for measuring photosynthesis in vitro for major tree species in northeastern China

doi:10.3724/SP.J.1258.2011.00452

Abstract

Abstract:

Aims Tree leaf photosynthesis is often difficult to measure in situ because of the physical inaccessibility of the tree canopy. One simple, inexpensive solution is to measure photosynthesis in vitro, but its validity and reliability for specific tree species need verification. Our goals were to determine (a) which in vitro measurement methods would result in detached leaves maintaining as high photosynthetic activity as in situ, (b) how long the photosynthetic activity of detached leaves can be sustained and (c) the reliability and feasibility of in vitro measurements.
Methods We selected seven major tree species with divergent ecophysiological characteristics in the temperate forest of northeastern China: coniferous (Pinus koraiensis and Larix olgensis), diffuse-porous (Betula platyphylla and Juglans mandshurica) and ring-porous (Fraxinus mandshurica, Ulmus macrocarpa and Quercus mongolica) species. We used the time for the photosynthetic rate to recover 90% of its pre-photoinducement value (T₉₀) and leaf transpiration rate (E) to assess the water supply and photosynthetic activity of detached leaves. Based on this, we determined an optimal in vitro protocol for measuring photosynthesis of detached leaves. We simultaneously monitored the duration of relatively stable photosynthetic rates after leaves had been detached. We then compared the differences in gas exchange parameters between in situ and in vitro measurements with the chosen protocol to determine the reliability of the protocol for each tree species.
Important findings The detached leaves of all measured tree species except for Q. mongolica had the potential to maintain relatively high, stable water supply and photosynthetic activity. The optimal protocol of in vitro photosynthesis measurements for the other six tree species was to insert the twigs or compound leaves into water immediately following detaching, girdle phloem about 3 cm from the cut and remove all leaves except the target ones. T₉₀differed significantly among the six tree species (p < 0.05), and the ring-porous tree species had significantly greater T₉₀than the other species. The leaf water supply and photosynthetic activity for the six species was effectively maintained for one hour following detachment, during which most of the gas exchange parameters measured did not differ significantly between in situ and in vitro measurements. Therefore, this study provided a feasible protocol of in vitro measurement of leaf photosynthesis for the six temperate tree species.

Key words: gas exchange, in situ, in vitro, photosynthetic activity, stable time, temperate forest

TANG Yan, WANG Chuan-Kuan. A feasible method for measuring photosynthesis in vitro for major tree species in northeastern China[J]. Chin J Plant Ecol, 2011, 35(4): 452-462.

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URL: https://www.plant-ecology.com/EN/10.3724/SP.J.1258.2011.00452

https://www.plant-ecology.com/EN/Y2011/V35/I4/452

Figures/Tables 6

Table 1 Protocols applied for in vitro measurements of leaf photosynthesis

随机区组设计 Randomized block design	处理 Treatment	处理描述 Treatment description	供试树种 Applied tree species
1	1	原位 in situ	BH、HY
	2	斜切细枝: 切口斜切的当年生细枝 Bevel twigs: current-year twigs were beveled from the cut	BH、HY
	3	环剥细枝: 切口处环剥去除3 cm左右韧皮部的当年生细枝 Girdle twigs: current-year twigs were girdled phloem about 3 cm from the cut	BH、HY
	4	环剥粗枝: 切口处环剥去除3 cm左右韧皮部的直径为2 cm左右的粗枝 Girdle branches: branches with diameter about 2 cm were girdled phloem about 3 cm from the cut	BH、HY
2	1	原位 in situ	HTQ、SQL
	3	环剥细枝 Girdle twigs	HTQ、SQL
	5	环剥复叶: 切口处环剥去除3 cm左右韧皮部的复叶 Girdle compound leaves: compound leaves were girdled phloem about 3 cm from the cut	HTQ、SQL

Fig. 1 Temporal dynamics of net photosynthetic rates (Pn) for the seven tree species with various treatments. BH, Betula platyphylla; HS, Pinus koraiensis; HTQ, Juglans mandshurica; HY, Ulmus macrocarpa; LYS, Larix olgensis; MGL, Quercus mongolica; SQL, Fraxinus mandshurica.

Fig. 2 Comparison of leaf transpiration rate (E) for the four tree species with various treatments (mean ± SE). BH, Betula platyphylla; HTQ, Juglans mandshurica; HY, Ulmus macrocarpa; SQL, Fraxinus mandshurica. Different lowercase letters mean significant differences among different treatments at p < 0.05 level.

Fig. 3 The time for the net photosynthetic rate recovering to 90% of its pre-photoinducement value (T90) and the difference in T90 between in situ and in vitro measurements (ΔT90) for the six tree species (mean ± SE). T90 for each species was the mean of T90 in various treatments (Table 1; Fig. 1); ΔT90 was the mean difference in T90 between in situ and various in vitro measurements. BH, Betula platyphylla; HS, Pinus koraiensis; HTQ, Juglans mandshurica; HY, Ulmus macrocarpa; LYS, Larix olgensis; SQL, Fraxinus mandshurica. Different lowercase letters mean significant differences among different treatments at p < 0.05 level.

Fig. 4 Temporal changes of the relative variation of net photosynthetic rates in vitro for the six tree species. BH, HY, HS and LYS use the method of girdling twigs, HTQ and SQL use the method of girdling compound leaves. BH, Betula platyphylla; HS, Pinus koraiensis; HTQ, Juglans mandshurica; HY, Ulmus macrocarpa; LYS, Larix olgensis; SQL, Fraxinus mandshurica.

Fig. 5 Comparison between in situ and in vitro measurements of light-saturated net photosynthetic rates (Pmax), apparent quantum yield (AQE), light saturation point (Ls), light compensation point (Lc), dark respiration rate (Rd), maximum rate of Rubisco carboxylation (Vcmax), maximum electron transport rate (Jmax), triose phosphate use rate (TPU), water use efficiency (WUE) and transpiration rate (E) for the six tree species (mean ± SE). BH, HY, HS and LYS use the method of girdling twigs, HTQ and SQL use the method of girdling detached compound leaves. *, difference in a specific parameter between in situ and in vitro measurements is significant (p < 0.05, two-tailed paired t-test) for a specific tree species. BH, Betula platyphylla; HS, Pinus koraiensis; HTQ, Juglans mandshurica; HY, Ulmus macrocarpa; LYS, Larix olgensis; SQL, Fraxinus mandshurica.

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