Mishra Kumud B, Mishra Anamika, Novotná Kateřina, Rapantová Barbora, Hodaňová Petra, Urban Otmar, Klem Karel
Global Change Research Institute, The Czech Academy of Sciences, v. v. i, Bělidla 986/4a, 603 00 Brno, Czech Republic.
Plant Methods. 2016 Nov 14;12:46. doi: 10.1186/s13007-016-0145-3. eCollection 2016.
Non-invasive and high-throughput monitoring of drought in plants from its initiation to visible symptoms is essential to quest drought tolerant varieties. Among the existing methods, chlorophyll fluorescence (ChlF) imaging has the potential to probe systematic changes in photosynthetic reactions; however, prerequisite of dark-adaptation limits its use for high-throughput screening.
To improve the throughput monitoring of plants, we have exploited their light-adaptive strategy, and investigated possibilities of measuring ChlF transients under low ambient irradiance. We found that the ChlF transients and associated parameters of two contrasting accessions, Rsch and Co, give almost similar information, when measured either after 20 min dark-adaptation or in the presence of - The fluorescence parameters, (Φ) and () resulting from this approach enabled us to differentiate accessions that is often not possible by well-established dark-adapted fluorescence parameter (/). Further, we screened ChlF transients in rosettes of well-watered and drought-stressed six accessions, under -, without any prior dark-adaptation. Relative water content (RWC) in leaves was also assayed and compared to the ChlF parameters. As expected, the RWC was significantly different in drought-stressed from that in well-watered plants in all the six investigated accessions on day-10 of induced drought; the maximum reduction in the RWC was obtained for Rsch (16%), whereas the minimum reduction was for Co (7%). Drought induced changes were reflected in several features of ChlF transients; combinatorial images obtained from pattern recognition algorithms, trained on pixels of image sequence, improved the contrast among drought-stressed accessions, and the derived images were well-correlated with their RWC.
We demonstrate here that ChlF transients and associated parameters measured even in the presence of low ambient irradiance preserved its features comparable to that of measured after dark-adaptation and discriminated the accessions having differential geographical origin; further, in combination with combinatorial image analysis tools, these data may be readily employed for early sensing and mapping effects of drought on plant's physiology via easy and fully non-invasive means.
从干旱开始到出现可见症状对植物干旱情况进行非侵入性和高通量监测对于寻找耐旱品种至关重要。在现有方法中,叶绿素荧光(ChlF)成像有潜力探测光合反应中的系统性变化;然而,暗适应的前提限制了其在高通量筛选中的应用。
为了提高对植物的通量监测,我们利用了它们的光适应策略,并研究了在低环境光照下测量ChlF瞬变的可能性。我们发现,当在约20分钟暗适应后或在存在-的情况下测量时,两个对比品种Rsch和Co的ChlF瞬变及相关参数给出了几乎相似的信息。这种方法得到的荧光参数(Φ)和()使我们能够区分品种,而这通常是通过成熟的暗适应荧光参数(/)无法实现的。此外,我们在没有任何预先暗适应的情况下,在-条件下筛选了六个品种的充分浇水和干旱胁迫莲座叶中的ChlF瞬变。还测定了叶片中的相对含水量(RWC)并与ChlF参数进行比较。正如预期的那样,在诱导干旱的第10天,所有六个被研究品种中,干旱胁迫下的RWC与充分浇水的植物相比有显著差异;Rsch的RWC降低最大(16%),而Co的降低最小(约7%)。干旱诱导的变化反映在ChlF瞬变的几个特征中;从基于图像序列像素训练的模式识别算法获得的组合图像提高了干旱胁迫品种之间的对比度,并且所得到的图像与它们的RWC有很好的相关性。
我们在此证明,即使在低环境光照下测量的ChlF瞬变及相关参数也保留了与暗适应后测量相当的特征,并区分了具有不同地理起源的品种;此外,结合组合图像分析工具,这些数据可以通过简单且完全非侵入性的方式很容易地用于早期感知和绘制干旱对植物生理的影响。