Prakash Jogadhenu Syama Sundara, Baig Masroor A, Bhagwat Anil S, Mohanty Prasanna
School of Life Sciences, Jawaharlal Nehru University, New Delhi, 110 067, India.
J Plant Physiol. 2003 Feb;160(2):175-84. doi: 10.1078/0176-1617-00529.
Structure and function of chloroplasts are known to after during senescence. The senescence-induced specific changes in light harvesting antenna of photosystem II (PSII) and photosystem I (PSI) were investigated in Cucumis cotyledons. Purified light harvesting complex II (LHCII) and photosystem I complex were isolated from 6-day non-senescing and 27-day senescing Cucumis cotyledons. The chlorophyll a/b ratio of LHCII obtained from 6-day-old control cotyledons and their absorption, chlorophyll a fluorescence emission and the circular dichroism (CD) spectral properties were comparable to the LHCII preparations from other plants such as pea and spinach. The purified LHCII obtained from 27-day senescing cotyledons had a Chl a/b ratio of 1.25 instead of 1.2 as with 6-day LHCII and also exhibited significant changes in the visible CD spectrum compared to that of 6-day LHCII, indicating some specific alterations in the organisation of chlorophylls of LHCII. The light harvesting antenna of photosystems are likely to be altered due to aging. The room temperature absorption spectrum of LHCII obtained from 27-day senescing cotyledons showed changes in the peak positions. Similarly, comparison of 77K chlorophyll a fluorescence emission characteristics of LHCII preparation from senescing cotyledons with that of control showed a small shift in the peak position and the alteration in the emission profile, which is suggestive of possible changes in energy transfer within LHCII chlorophylls. Further, the salt induced aggregation of LHCII samples was lower, resulting in lower yields of LHCII from 27-day cotyledons than from normal cotyledons. Moreover, the PSI preparations of 6-day cotyledons showed Chl a/b ratios of 5 to 5.5, where as the PSI sample of 27-day cotyledons had a Chl a/b ratio of 2.9 suggesting LHCII association with PSI. The absorption, fluorescence emission and visible CD spectral measurements as well as the polypeptide profiles of 27-day cotyledon-PSI complexes indicated age-induced association of LHCII of PSII with PSI obtained from 27-day cotyledons. We modified our isolation protocols by increasing the duration of detergent Triton X-100 treatment for preparing the PSI and LHCII complexes from 27-day cotyledons. However, the PSI complexes isolated from senescing samples invariably proved to have significantly low Chl a/b ratio suggesting an age induced lateral movement and possible association of LHCII with PSI complexes. The analyses of polypeptide compositions of LHCII and PSI holocomplexes isolated from 6-day control and 27-day senescing cotyledons showed distinctive differences in their profiles. The presence of 26-28 kDa polypeptide in PSI complexes from 27-day cotyledons, but not in 6-day control PSI complexes is in agreement with the notion that senescence induced migration of LHCII to stroma lamellae and its possible association with PSI. We suggest that the migration of LHCII to the stroma lamellae region and its possible association with PSI might cause the destacking and flattening of grana structure during senescence of the chloroplasts. Such structural changes in light harvesting antenna are likely to alter energy transfer between two photosystems. The nature of aging induced migration and association of LHCII with PSI and its existence in other senescing systems need to be estimated in the future.
已知叶绿体的结构和功能在衰老过程中会发生变化。我们研究了黄瓜子叶中光系统II(PSII)和光系统I(PSI)的光捕获天线在衰老诱导下的特定变化。从6天龄未衰老和27天龄衰老的黄瓜子叶中分离出纯化的光捕获复合体II(LHCII)和光系统I复合体。从6天龄对照子叶中获得的LHCII的叶绿素a/b比值及其吸收、叶绿素a荧光发射和圆二色性(CD)光谱特性与从豌豆和菠菜等其他植物中制备的LHCII相当。从27天龄衰老子叶中获得的纯化LHCII的Chl a/b比值为1.25,而6天龄LHCII的该比值为1.2,并且与6天龄LHCII相比,其可见CD光谱也表现出显著变化,这表明LHCII中叶绿素的组织发生了一些特定改变。光系统的光捕获天线可能会因衰老而改变。从27天龄衰老子叶中获得的LHCII的室温吸收光谱在峰值位置上发生了变化。同样,将衰老子叶中LHCII制剂的77K叶绿素a荧光发射特性与对照进行比较,发现峰值位置有小的偏移且发射轮廓发生改变,这表明LHCII叶绿素内的能量转移可能发生了变化。此外,盐诱导的LHCII样品聚集较低,导致从27天龄子叶中获得的LHCII产量低于正常子叶。而且,6天龄子叶的PSI制剂的Chl a/b比值为5至5.5,而27天龄子叶的PSI样品的Chl a/b比值为2.9,这表明LHCII与PSI相关联。27天龄子叶-PSI复合体的吸收、荧光发射和可见CD光谱测量以及多肽谱表明,衰老诱导PSII的LHCII与从27天龄子叶中获得的PSI发生关联。我们通过增加去污剂Triton X-100处理的持续时间来修改我们的分离方案,以从27天龄子叶中制备PSI和LHCII复合体。然而,从衰老样品中分离出的PSI复合体总是被证明具有显著较低的Chl a/b比值,这表明衰老诱导了LHCII的横向移动以及其与PSI复合体的可能关联。对从6天龄对照和27天龄衰老子叶中分离出的LHCII和PSI全复合体的多肽组成分析显示,它们的谱图存在明显差异。27天龄子叶的PSI复合体中存在26 - 28 kDa多肽,而6天龄对照PSI复合体中不存在,这与衰老诱导LHCII迁移到基质类囊体及其与PSI可能关联的观点一致。我们认为,LHCII向基质类囊体区域的迁移及其与PSI的可能关联可能导致叶绿体衰老过程中基粒结构的解堆叠和扁平化。光捕获天线的这种结构变化可能会改变两个光系统之间的能量转移。衰老诱导的LHCII与PSI的迁移和关联的性质及其在其他衰老系统中的存在情况,未来需要进行评估。