东北主要树种光合作用可行的离体测定方法

doi:10.3724/SP.J.1258.2011.00452

摘要/Abstract

摘要：

树木光合作用的测定常因植株高大而难以开展, 其中离体测定是解决途径之一。但离体测定的方法及其可靠性因树种而异。选取东北东部温带森林中特性各异的7种主要树种: 针叶树(红松(Pinus koraiensis)、长白落叶松(Larix olgensis))、散孔材(白桦(Betula platyphylla)、胡桃楸(Juglans mandshurica))和环孔材(水曲柳(Fraxinus mandshurica)、黄榆(Ulmus macrocarpa)、蒙古栎(Quercus mongolica)), 首先采用光合速率恢复到光合诱导前稳定值90%的时间(T₉₀)长短和叶片蒸腾速率(E)的大小评估离体叶片水分供应状况及其光合活力, 以此确定较优的离体测定方案; 同时, 观测离体叶片的光合活力稳定时间; 最后通过比较原位测定和采用所确定的较优离体方案测定的各树种叶片气体交换参数, 论证采用离体测定光合作用的可靠性。结果表明: 除蒙古栎外的6个树种的离体叶片均具有较高、较稳定的水分供应和光合活力。离体枝条或复叶插入水中, 环剥去除切口处3 cm左右的韧皮部和剩余叶片的方法, 是这6个温带树种叶片光合能力的较优离体测定方法。6个树种叶片的T₉₀受树木特性的影响而差异显著(p < 0.05), 其中环孔材树种的T₉₀显著高于散孔材和针叶树种。6个树种离体叶片在1 h内均有较高、较稳定的水分供应和光合活力。在此期间离体所测得的绝大多数叶片的气体交换参数与其原位测定值之间的差异不显著。该研究提出了可行的树木叶片光合作用的离体测定方案, 适用于蒙古栎以外的其他6个温带树种。

关键词: 气体交换, 原位, 离体, 光合活力, 温带森林, 稳定时间

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

唐艳, 王传宽. 东北主要树种光合作用可行的离体测定方法. 植物生态学报, 2011, 35(4): 452-462. DOI: 10.3724/SP.J.1258.2011.00452

TANG Yan, WANG Chuan-Kuan. A feasible method for measuring photosynthesis in vitro for major tree species in northeastern China. Chinese Journal of Plant Ecology, 2011, 35(4): 452-462. DOI: 10.3724/SP.J.1258.2011.00452

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

图/表 6

表1 供试叶片光合作用的离体测定方案

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

图1 7个树种不同处理下净光合速率的时间进程。BH, 白桦; HS, 红松; HTQ, 胡桃楸; HY, 黄榆; LYS, 长白落叶松; MGL, 蒙古栎; SQL,水曲柳。

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.

图2 比较不同处理下4个树种的叶片蒸腾速率(E) (平均值±标准误差)。BH, 白桦; HTQ, 胡桃楸; HY, 黄榆; SQL, 水曲柳。不同小写字母表示处理间差异显著(p < 0.05)。

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.

图3 6个树种净光合速率恢复到光合诱导前稳定值90%的时间(T90)及原位和离体的T90之差(ΔT90 = T90离体-T90原位) (平均值±标准误差)。各树种的T90是不同处理方法下(表1; 图1) T90的平均值; ΔT90是原位和不同离体处理方法的T90之差的平均值。BH, 白桦; HS, 红松; HTQ, 胡桃楸; HY, 黄榆; LYS, 长白落叶松; SQL,水曲柳。不同小写字母表示处理间差异显著(p < 0.05)。

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.

图4 离体条件下6个树种净光合速率相对变异(RV)的时间变化。BH、HY、HS和LYS采用环剥细枝的离体方法, HTQ和SQL采用环剥复叶的离体方法。BH, 白桦; HS, 红松; HTQ, 胡桃楸; HY, 黄榆; LYS, 长白落叶松; SQL,水曲柳。

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.

图5 6个树种的最大净光合速率(Pmax)、表观量子效率(AQE)、光饱和点(Ls)、光补偿点(Lc)、暗呼吸速率(Rd)、最大羧化速率(Vcmax)、最大电子传递速率(Jmax)、磷酸丙糖利用速率(TPU)、水分利用效率(WUE)和蒸腾速率(E)原位测定值与离体测定值之间的比较(平均值±标准误差)。BH、HY、HS和LYS采用环剥细枝的离体方法, HTQ和SQL采用环剥复叶的离体方法。*, 某个树种叶片的某个参数的原位测定值和离体测定值之间差异显著(p < 0.05, 双侧成对t检验)。BH, 白桦; HS, 红松; HTQ, 胡桃楸; HY, 黄榆; LYS, 长白落叶松; SQL, 水曲柳。

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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