In order to further understand the inner mechanism of plant respiration and the relationship between respiration and photosynthesis of plant under light, our objective was to reveal the relationship between cyanide-resistant respiration and photosynthesis in bean (Phaseolus vulgaris) leaves under light.
Methods
By exposing the dark-grown leaves to light for 10 h, changes in total respiration (Vt), the cyanide-resistant respiration (Valt), photosynthetic O2 evolution, and photosynthetic CO2 fixation, and the effects of inhibitor of the cyanide-resistant respiration on photosynthesis were measured and analyzed. We also measured and analyzed changes in the ratio of cyanide-resistant respiration, total respiration, and photosynthetic CO2 fixation when leaves in the dark were exposed to a brief period (10 min) of light. Important findings After exposing the dark-grown leaves to light for 10 h, Vt, Valt, and the value of Valt/Vt all increased. During the process, time-course analysis after the onset of illumination demonstrated that the induction of the cyanide-resistant respiration in light was prior to the formations of the photosynthetic O2 evolution and CO2 fixation. This observation indicates that the induction of the cyanide-resistant respiration by light is independent of photosynthesis. The dark-grown leaves pretreated with 1 mmol·L-1 salicylhydroxamic acid (SHAM; the inhibitor of the cyanide-resistant respiration) were exposed to illumination for 10 h, and SHAM did not result in apparent modification of the photosynthetic O2 evolution and CO2 fixation in the leaves when exposed to light. This observation also showed that there was no direct linkage between photosynthesis and the cyanide-resistant respiration when the dark-grown leaves were exposed to light. In addition, under the condition of darkness, 10 min of light illumination obviously increased the value of Valt/Vt but did not significantly affect the level of photosynthetic CO2 fixation.
Results
indicate that the induction of the cyanide-resistant respiration by light may be independent of photosynthesis. Light should have a direct influence on induction of the cyanide-resistant respiration.
FENGHan-Qing, GUANDong-Dong, JIAOQing-Song, JIALing-Yun, SUNKun. Analysis of the relationship between cyanide-resistant respiration and photosynthesis under light in Phaseolus vulgaris leaves. Chinese Journal of Plant Ecology, 2015, 39(1): 104-109 https://doi.org/10.17521/cjpe.2015.0011
总RNA使用Trizol法提取。以烟草中交替氧化酶编码基因的cDNA片段(Whelan et al., 1995)为探针, 利用ECL非放射性核酸标记试剂盒(Enzo Diagnostics, Little Chalfont, UK)对其进行标记和Northern杂交实验, 探针标记和信号检测等过程依据试剂盒所提供的实验指导说明进行。
Fig. 1 Changes in total respiration (Vt), capacity of cyanide- resistant respiration (Valt) and the ratio of Valt to Vt in dark- grown leaves exposed to continuous light for 10 h. These are individual samples taken during four different experiments. Time indicates hours after starting illumination. Values are mean values ± SD of four independent experiments. The horizontal axis shows the time after exposing to light. Valt/Vt was computed based on the average values of Vt and Valt.
Fig. 3 Changes in oxygen evolution (■) and carbon dioxide fixation (□) in whole chloroplasts of the dark-grown leaves exposed to 10 h of continuous light. Values are mean values ± SD of four independent experiments. The horizontal axis shows the time after exposing to light.
2.3 黑暗中生长的叶片转至光照过程中SHAM对光合CO2固定和光合放氧的影响
SHAM是常用的抗氰呼吸的化学抑制剂(Chivasa et al., 1997; Chivasa & Carr, 1998)。将黑暗下生长的叶片在转至光照前用1 mmol·L-1的SHAM (所用浓度已被报道能有效抑制抗氰呼吸的活性, Bartoli et al., 2005)处理叶片以抑制其抗氰呼吸的活性。将用SHAM处理的叶片和未经过SHAM处理的黑暗下生长的叶片置于光照下照射10 h并对比二者的光合CO2固定和放氧水平, 发现SHAM的处理并没有显著性地影响植物叶片光合CO2固定和放氧速率(表1)。
Table 1
表1
表1 黑暗中生长的叶片转至光照过程中水杨基氧肟酸(SHAM)对叶片光合CO2固定和放氧速率的影响
Table 1 The effects of salicylhydroxamic acid (SHAM) on photosynthetic oxygen evolution rate and carbon dioxide fixation rate when the dark-grown leaves were exposed to continuous light
光照的时间 Time of illumination (h)
SHAM对于光合作用的影响 Effects of SHAM on photosynthesis (% of control)
光合放氧 Oxygen evolution
光合CO2固定 CO2 fixation
0
100a
100a
2
100a
100a
4
98 ± 5a
100a
6
101 ± 3a
100a
8
97 ± 4a
96 ± 4a
10
102 ± 4a
95 ± 3a
The leaves without being subjected to SHAM treatment were assigned the values of control (100%). Values are mean values ± SD of four independent experiments. Time indicates hours following commencement of illumination. Same letters indicate there are no significant differences between the SHAM treatment and the control (p < 0.05).以未经过SHAM处理的叶片作为对照(100%)。数值为4次独立实验的平均值±标准偏差。所示时间为叶片转至光照下的时间。相同字母表示SHAM处理和对照之间无显著性差异(p < 0.05)。
Fig. 4 Effect of short period of illumination on the ratio of cyanide-resistant respiration pathway to total respiration (Valt/Vt, %) (■) and photosynthetic CO2 fixation rate (□). One-week-old plants grown in 12 h light/12 h dark photoperiods were transferred to darkness and then received 10 min plus of light once every 12 h. Leaves in darkness were assigned the values of control (100%). Time indicates hours in darkness. Results are mean values ± SD of four independent experiments.
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Restriction of mitochondrial oxidative metabolism leads to suppression of photosynthetic carbon assimilation but not of photochemical electron transport in pea mesophyll protoplasts
SiedowJN, DayDA (2000). Respiration and photorespiration. In: Buchanan B, Gruissem W, Jones R eds. Biochemistry and Molecular Biology of Plants. American Society of Plant Physiologists, Rockville, USA. 676-728.
Mitochondrial alternative oxidase pathway protects plants against photoinhibition by alleviating inhibition of the repair of photodamaged PSII through preventing formation of reactive oxygen species in Rumex K-1 leaves
... 以前的研究表明, 光能够通过影响光受体的表达而直接影响线粒体的电子传递链(Liscum et al., 2003; Escobar et al., 2004).而最近的研究发现, 光照可以通过刺激光敏色素、隐花色素和向光素等光受体而诱导拟南芥(Arabidopsis thaliana)交替氧化酶基因的表达(Zhang et al., 2010; Xu et al., 2011).这些研究均提示了光作为一种直接的信号去调节植物抗氰呼吸的水平.因此, 我们推测: 在本研究中光对于抗氰呼吸的诱导作用可能是由于光照刺激了这些光受体从而导致了交替氧化酶表达水平上升所致.以前的研究已经揭示从黑暗转为光照会导致植物活性氧水平的上升(Kim et al., 2008); 因此光照下抗氰呼吸的上升可能通过减少这些活性氧的产生而对植物具有保护作用.此外, 也有研究发现从黑暗转为光照会导致植物生物合成水平的增加(Bruick & Mayfield, 1999). 而糖酵解和三羧酸循环过程中产生的诸多碳物质是植物进行生物合成反应所需的重要前体; 而抗氰呼吸被认为能够促进糖酵解和三羧酸循环的快速运转(Mackenzie & McIntosh, 1999), 因此, 本文推测光照下抗氰呼吸的上升也可能在一定程度上和植物的生物合成反应有关. ...
Cyanide restores N gene-mediated resistance to tobacco mosaic virus in transgenic tobacco expressing salicylic acid hydroxylase
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1998
... SHAM是常用的抗氰呼吸的化学抑制剂(Chivasa et al., 1997; Chivasa & Carr, 1998).将黑暗下生长的叶片在转至光照前用1 mmol·L-1的SHAM (所用浓度已被报道能有效抑制抗氰呼吸的活性, Bartoli et al., 2005)处理叶片以抑制其抗氰呼吸的活性.将用SHAM处理的叶片和未经过SHAM处理的黑暗下生长的叶片置于光照下照射10 h并对比二者的光合CO2固定和放氧水平, 发现SHAM的处理并没有显著性地影响植物叶片光合CO2固定和放氧速率(表1). ...
Salicylic acid interferes with tobacco mosaic virus replication via a novel salicylhydroxamic acid-sensitive mechanism
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1997
... SHAM是常用的抗氰呼吸的化学抑制剂(Chivasa et al., 1997; Chivasa & Carr, 1998).将黑暗下生长的叶片在转至光照前用1 mmol·L-1的SHAM (所用浓度已被报道能有效抑制抗氰呼吸的活性, Bartoli et al., 2005)处理叶片以抑制其抗氰呼吸的活性.将用SHAM处理的叶片和未经过SHAM处理的黑暗下生长的叶片置于光照下照射10 h并对比二者的光合CO2固定和放氧水平, 发现SHAM的处理并没有显著性地影响植物叶片光合CO2固定和放氧速率(表1). ...
Light regulation of the Arabidopsis respiratory chain. Multiple discrete photo- receptor responses contribute to induction of type II NAD(P)H dehydrogenase genes
1
2004
... 以前的研究表明, 光能够通过影响光受体的表达而直接影响线粒体的电子传递链(Liscum et al., 2003; Escobar et al., 2004).而最近的研究发现, 光照可以通过刺激光敏色素、隐花色素和向光素等光受体而诱导拟南芥(Arabidopsis thaliana)交替氧化酶基因的表达(Zhang et al., 2010; Xu et al., 2011).这些研究均提示了光作为一种直接的信号去调节植物抗氰呼吸的水平.因此, 我们推测: 在本研究中光对于抗氰呼吸的诱导作用可能是由于光照刺激了这些光受体从而导致了交替氧化酶表达水平上升所致.以前的研究已经揭示从黑暗转为光照会导致植物活性氧水平的上升(Kim et al., 2008); 因此光照下抗氰呼吸的上升可能通过减少这些活性氧的产生而对植物具有保护作用.此外, 也有研究发现从黑暗转为光照会导致植物生物合成水平的增加(Bruick & Mayfield, 1999). 而糖酵解和三羧酸循环过程中产生的诸多碳物质是植物进行生物合成反应所需的重要前体; 而抗氰呼吸被认为能够促进糖酵解和三羧酸循环的快速运转(Mackenzie & McIntosh, 1999), 因此, 本文推测光照下抗氰呼吸的上升也可能在一定程度上和植物的生物合成反应有关. ...
Differential expression of the multigene family encoding the soybean mitochondrial alternative oxidase
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1997
... 光是重要的生态学因子, 是植物进行光合作用的原初能量来源.而呼吸作用是植物能量和物质代谢的中心环节.因此, 光合作用和呼吸作用之间的关系始终是植物生态学和植物生理学关注的重要问题.一些研究已经发现光照能够导致植物抗氰呼吸水平的上升(Obenland et al., 1990; Finnegan et al., 1997; Ribas-carbo et al., 2000).目前的主流观点认为, 抗氰呼吸水平在光照下的上升主要与植物的光合作用有关, 即在光照下叶绿体中的部分还原力可以通过苹果酸/草酰乙酸穿梭机制运输到线粒体中, 并主要通过抗氰呼吸途径进行氧化, 因而增加了抗氰呼吸的水平(Padmasree et al., 2002; Yoshida et al., 2007).除此之外, 叶绿体在光合作用过程中所产生的乙醇酸会通过光呼吸路径在线粒体中形成甘氨酸, 而这些甘氨酸也能刺激抗氰呼吸途径的运行(Igamberdiev et al., 1997).同时, 抗氰呼吸水平在光照下的上升也被认为是帮助叶绿体去消耗光合作用过程中产生的过多的还原力, 从而避免叶绿体处于过度还原的状态(Svensson & Rasmusson, 2001; Padmasree et al., 2002; Raghavendra & Padmasree, 2003; Bartoli et al., 2005). ...
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Restriction of mitochondrial oxidative metabolism leads to suppression of photosynthetic carbon assimilation but not of photochemical electron transport in pea mesophyll protoplasts
Mitochondrial alternative oxidase pathway protects plants against photoinhibition by alleviating inhibition of the repair of photodamaged PSII through preventing formation of reactive oxygen species in Rumex K-1 leaves
2
2011
... 以往的研究发现: 当用抑制剂等方法抑制抗氰呼吸在光照下的上升, 会导致植物光合CO2固定和放氧速率水平的下降(Padmasree & Raghavendra, 2001; Yoshida et al., 2006; Zhang et al., 2011, 2012), 这也是光合作用和抗氰呼吸具有偶联关系的重要证据.有学者认为, 这是由于光照会导致叶片中还原力的积累, 而被积累的还原力会使得叶绿体中的氧分子被还原为活性氧; 由于高度的反应活性, 活性氧会引起叶绿体光合机构的氧化损伤, 从而引起植物光合作用的下降.而抗氰呼吸水平在光照下的上升有助于叶绿体消耗光合作用过程中产生的过多的还原力, 从而保护了光合机构免受活性氧的损伤(Yoshida et al., 2006, 2007; Zhang et al., 2011, 2012).而本研究发现, 将黑暗下生长的植物转至光照前用SHAM处理叶片并不会导致光照下叶片光合CO2固定和放氧速率的显著性变化(表1), 表明了在黑暗下生长的叶片在转至10 h的光照过程中抗氰呼吸并未与光合作用产生偶联. ...
, 管东东, 焦青松, 贾凌云, 孙坤
