叶片尺度光合荧光实验设计

1、叶片尺度光合叶片尺度光合/荧荧光光实验设计实验设计 ecotek 不同时间尺度上的实验不同时间尺度上的实验 1 季节尺度动态变化 2 日尺度动态变化 3 小时尺度动态变化 4 分钟尺度动态变化 5 秒尺度动态变化 1 季节尺度动态变化季节尺度动态变化 1.1 光合参数季节动态光合参数季节动态 1.2 荧光参数季节动态荧光参数季节动态 1.1 光合参数季节动态光合参数季节动态 (david a. ramrez et al. 2012) 光合参数季节动态解释的问题光合参数季节动态解释的问题 v 不同种植物对降水格局变化的响应不同种植物对降水格局变化的响应 the relationship betw

2、een water availability, and rgr and carbon assimilation in c. ladanifer , and the lack of any relationship in e. arborea suggest that the former has an enhanced capacity to harness unpredictable rainfall pulses compared with the latter. v 改变群落中植物种的优势地位改变群落中植物种的优势地位 these contrasting responses to wat

3、er availability indicate that the projected changes in rainfall with global warming could alter the competitive ability of these two species, and contribute to changes in plant dominance in mediterranean shrublands. (david a. ramrez et al. 2012) 光合参数季节动态实验方法光合参数季节动态实验方法 v 测量时间：测量时间： net carbon assim

4、ilation (a) and stomatal conductance (gs) were measured early in the morning (08:00 hours solar time) to avoid midday photo inhibition. v 环境条件的控制：环境条件的控制： chamber conditions set to match typical meteorological conditions in the early morning during the growing season: photosynthetically active radia

5、tion at 1000mmol m-2s-1, reference co2 concentration at 400ppm, leaf temperature at 25, and vapour pressure deficit at 1.5-2.0kpa. (david a. ramrez et al. 2012) 光合参数季节动态实验小结光合参数季节动态实验小结 v 实验仪器：实验仪器：li-6400/xt ； 6400-40 v 采样：略采样：略 v 时间：定时时间：定时 v 重复数：重复数：n=8 v 光：控制光：控制 v 温度：控制温度：控制 v 二氧化碳浓度：控制二氧化碳浓度：控

6、制 v 湿度湿度: 维持在一定范围内维持在一定范围内 (david a. ramrez et al. 2012) 荧光参数季节动态荧光参数季节动态 (binghua liu et al. 2012) 荧光参数季节动态解释的问题荧光参数季节动态解释的问题 v砧木的不同导致对干旱的响应不同砧木的不同导致对干旱的响应不同 our present study revealed drought-related reductions in chl content, pn and fv/fm the extent of which were more notable in plants on m. hupe

7、hensis.thus, the growth and biomass production of apple trees in response to drought depends upon the choice of rootstock. (binghua liu et al. 2012) 荧光参数季节动态实验方法荧光参数季节动态实验方法 v 仪器仪器 chlorophyll fluorescence was measured with an integrating fluorescence fluorometer (6400-40 leaf chamber fluorometer; l

8、i-cor). v fo的测量的测量 after the samples were adapted to darkness for 1 h, the minimum fluorescence (fo) was measured with weak modulated irradiation (7,000 molm-2s-1) was applied to determine the maximum chlorophyll fluorescence yield (fm) and maximum photochemical efficiency(fv/fm). (binghua liu et al

9、. 2012) 荧光参数季节动态小结荧光参数季节动态小结 v 实验仪器：实验仪器：li-6400/xt ；6400-40 v 样品：文中没有提及，但是应该采用相同的标准；样品：文中没有提及，但是应该采用相同的标准； v 时间：定时；时间：定时； v 重复数：重复数：n=6； v 光：测量光光：测量光7,000 mol m-2s-1 ； v 温度：没有提及温度：没有提及 v 二氧化碳浓度：没有提及二氧化碳浓度：没有提及 v 湿度湿度: 没有提及没有提及 (binghua liu et al. 2012) 2 日尺度动态变化日尺度动态变化 v2.1 光合参数日动态光合参数日动态 v2.2 荧光参数

10、日动态荧光参数日动态 光合光合/荧光参数日动态荧光参数日动态 （孙东宝，王庆锁，（孙东宝，王庆锁，2012） 光合光合/荧光参数日动态解释的问题荧光参数日动态解释的问题 v灌水改变了苜蓿叶片灌水改变了苜蓿叶片pn的日变化格局。的日变化格局。 v灌水较多的处理灌水较多的处理(w3 和和w2), 苜蓿叶片没有出现苜蓿叶片没有出现 光合光合“ 午休午休” 现象现象, pn 的日变化趋势呈现的日变化趋势呈现 “ 单峰单峰” 型。型。 v而灌水较少和不灌水的处理而灌水较少和不灌水的处理(w1 和和w0), 苜蓿叶苜蓿叶 片出现了明显的光合片出现了明显的光合“午休午休” 现象现象, 其其pn的日的日 变化

11、进程呈现变化进程呈现“双峰双峰”型。型。 （孙东宝（孙东宝 subsequently, a/par curves were obtained at 40pa co2 partial pressure. v 水分利用效率、光补偿点、光饱和点的计算水分利用效率、光补偿点、光饱和点的计算 light use efficiencies (lue), light compensating points (lcp) and light saturation points (lsp) were determined from these curves. non-linear regression techn

12、iques for estimating these parameters were followed from ogren and evans (1993). （paulo et al. 2012) 光响应曲线实验小结光响应曲线实验小结 v 实验仪器：实验仪器：li-6400（xt）6400-40 v 采样：略采样：略 v 时间：根据设置的光强梯度，大致估算出实验耗时。选择叶片标准时间：根据设置的光强梯度，大致估算出实验耗时。选择叶片标准 要一致。要一致。 v 重复数：重复数：n=5 v 光适应：没做光适应：没做 ；改用的是低二氧化碳浓度适应改用的是低二氧化碳浓度适应。 v 温度：控制温度：

13、控制 v 二氧化碳浓度：控制二氧化碳浓度：控制 v 湿度湿度: 略略 温度响应曲线温度响应曲线 (masabumi et al. 2012) 温度响应曲线解释的问题温度响应曲线解释的问题 v 温度与二氧化碳的协同作用温度与二氧化碳的协同作用 photosynthetic carbon assimilation rates were synergistically enhanced above 30 under elevated co2 because of the higher etr and suppression of photorespiration under elevated co2

14、 (fig. 2a, farquhar et al. 1980). v 高温下，二氧化碳对光合的促进现象高温下，二氧化碳对光合的促进现象 it is noteworthy that based on the temperature-dependent photosynthesis, higher photosynthetic rates were observed above 30 in elevated-co2 -grown plants measured under elevated co2than in ambient-grown ones measured under ambient

15、co2. v 结论结论 this suggests that elevated co2 would boost photosynthetic carbon assimilation at higher temperatures even though photosynthetic down regulation occurred. (masabumi et al. 2012) 温度响应曲线实验方法温度响应曲线实验方法 v the temperature-dependent net photosynthetic rate (pn), quantum yield of psii electron

16、transport ( psii), photochemical quenching (qp),photochemical efficiency of the open psii (fv/f m) and quantum yield of non-regulated y(no) and regulated non-photochemical energy loss in psii y(npq) were measured at a photosynthetic steady state, at various temperatures （15, 20, 25, 30, 35 and 40）,

17、ambient co2 concentrations of 400 and 800mol mol 1 and saturating pfd of 1000 mol m2s1. (masabumi et al. 2012) 温度响应曲线实验小结温度响应曲线实验小结 v 实验仪器：实验仪器：li-6400（xt）6400-40 v 采样：采样： 生长生长50天完全展开的叶片天完全展开的叶片 v 时间：根据设置的温度梯度，大致估算出实验耗时。时间：根据设置的温度梯度，大致估算出实验耗时。 v 重复数：重复数：n=8 v 光适应：光适应：30分钟分钟 v 温度：适应某个温度至少温度：适应某个温度至少3

18、0分钟分钟 v 二氧化碳浓度：控制二氧化碳浓度：控制 v 湿度湿度: 控制（详见原文）。控制（详见原文）。 (masabumi et al. 2012) 湿度响应曲线湿度响应曲线 （顾礼力等，（顾礼力等，2012） 湿度响应曲线解释的问题湿度响应曲线解释的问题 v设施栽培中需要精细化管理，使大棚中保持适设施栽培中需要精细化管理，使大棚中保持适 宜的温湿度，从而提高杨梅净光合速率。宜的温湿度，从而提高杨梅净光合速率。 湿度响应曲线测量方法湿度响应曲线测量方法 v为了检验其关系，实验控制光强为了检验其关系，实验控制光强 1500molm-2s-1，co2 浓度浓度375 molmol-1， 温度温

19、度32 ，采取，采取50% 、55% 、60% 、65% 、 70% 、75% 、80% 共共7 个湿度梯度测定杨梅净个湿度梯度测定杨梅净 光合速率。光合速率。 湿度响应曲线实验小结湿度响应曲线实验小结 v 实验仪器：实验仪器：li-6400（xt）6400-40/6400-02b/6400-18 v 采样：略采样：略 v 时间：略时间：略 v 重复数：略重复数：略 v 光适应：光适应：30分钟分钟 v 温度：略温度：略 v 二氧化碳浓度：控制二氧化碳浓度：控制 v 湿度湿度:设置湿度梯度设置湿度梯度 二氧化碳响应曲线二氧化碳响应曲线 (masabumi et al. 2012) 二氧化碳响应

20、曲线解释的问题二氧化碳响应曲线解释的问题 v 生在在高二氧化碳浓度下的植物生在在高二氧化碳浓度下的植物rubisco羧化能力下降羧化能力下降 plants grown under elevated co2 exhibited photosynthetic down regulation, indicated by a decrease in the carboxylation capacity of rubisco. v 白桦树幼苗最大羧化速率和白桦树幼苗最大羧化速率和rubp再生速率下降再生速率下降 white birch seedlings grown at elevated co2 un

21、der limited n supply showed photosynthetic down regulation, indicated by decreases in the maximum rates of rubp carboxylation (vcmax) and rubp regeneration(jmax ) (masabumi et al. 2012) 二氧化碳响应曲线测量方法二氧化碳响应曲线测量方法 v the maximum rates of rubp carboxylation (vcmax)and rubp regeneration were derived from

22、photosynthetic responses to various ci at a leaf temperature of 25 and saturating photon flux density (pfd) of 1000mol m2s1, which was provided by a red/blue light emitting diode (led) array （6400-40;licor） with blue light comprising 10% of total pfd . (masabumi et al. 2012) 二氧化碳响应曲线实验小结二氧化碳响应曲线实验小结

23、 v 实验仪器：实验仪器：li-6400（xt）6400-40 v 采样：生长采样：生长50天完全展开的叶片天完全展开的叶片 v 时间：依据二氧化碳浓度梯度个数而定时间：依据二氧化碳浓度梯度个数而定 v 重复数：重复数：n=8 v 光：适应光：适应30分钟分钟 v 温度：适应温度：适应30分钟分钟 v 二氧化碳浓度：梯度二氧化碳浓度：梯度 v 湿度湿度:不控不控 荧光荧光-光响应曲线光响应曲线 (mitsutoshi et al. 2012) 荧光荧光-光响应曲线解释的问题光响应曲线解释的问题 v the shade leaves of the japanese oak grown withi

24、n a crown were suggested to adjust their n investment to maintain higher photosynthetic capacities compared with those required to maximize the net carbon gain, which may facilitate the dissipation of the excessive light energy of sunflecks to circumvent photoinhibition in cooperation with thermal e

25、nergy dissipation. (mitsutoshi et al. 2012) 荧光荧光-光响应曲线测量方法光响应曲线测量方法 v measurements of gas exchange and chlorophyll fluorescence were conducted on 17 mature leaves of the japanese oak with a portable photosynthesis measuring system (li-6400, licor) combined with a leaf chamber fluorometer (6400-40, l

26、icor) in mid-july. v the net photosynthetic rate (pn), quantum yield of psii elec- tron transport ( psii), photochemical quenching (qp)and photochemical efficiency of the open psii (fv/fm)were measured at a photosynthetic steady state, under an ambient co2 concentration of 360molmol1and at various p

27、pfd (0, 50, 100, 200, 300, 600, 1000,1500 and 2000 mol m2s1) which was provided by a red/blue led array (640040, licor) with blue light comprising 10 % of total ppfd. (mitsutoshi et al. 2012) 荧光荧光-光响应曲线实验小结光响应曲线实验小结 v 实验仪器：实验仪器：li-6400（xt）6400-40 v 采样：成熟叶片采样：成熟叶片 v 时间：依据光强梯度个数而定时间：依据光强梯度个数而定 v 重复数：略

28、重复数：略 v 光：适应光：适应30分钟分钟 v 温度：控制温度：控制 v 二氧化碳浓度：控制二氧化碳浓度：控制 v 湿度湿度:略略 4 分钟尺度动态变化分钟尺度动态变化 4.1 光合参数分钟动态光合参数分钟动态（auto log） 4.2 荧光诱导曲线（荧光诱导曲线（fluorescence induction curve） 4.3 暗驰豫动力学（暗驰豫动力学（relaxation of the kautsky curve） 分钟尺度动态变化分钟尺度动态变化 (veronica et al. 2012) 分钟尺度动态变化分钟尺度动态变化解释的问题解释的问题 v genes nhx1 and

29、nhx2 were highly expressed in guard cells, and stomatal function was defective in mutant plants, further compromising their ability to regulate water relations. v together, these results show that tonoplast- localized nhx proteins are essential for active k+ uptake at the tonoplast, for turgor regul

30、ation, and for stomatal function. (veronica et al. 2012) 光合参数光合参数分钟尺度分钟尺度变化测量方法变化测量方法 v leaf gas exchange was determined using the open gas exchange system li-6400 (li-cor) equipped with the chamber head (li-6400-40;li-cor) that allowed full control of light, co2, and humidity. v stomatal conductanc

31、e (gs; mmol m-2s-1) and the net photosynthetic rate (an; mmol m-2s-1) were measured in 3-week-old plants of the wild type and mutant line l2 grown hydroponically in lak standard solution. (veronica et al. 2012) 光合参数光合参数分钟尺度分钟尺度变化测量方法变化测量方法 v leaf responses to osmotic shock were recorded in fully exp

32、anded leaves, attached to the plant, under ambient co2 and a saturating ppfd of 150mmol m- 2s-1 leaves were allowed to equilibrate under those conditions for at least 10 min and then subjected to osmotic shock by 20% peg6000 in lak medium. v measurements were recorded every 30 s over a period of 120

33、 min. a total of 12 measurements for each genotype (six plants per line and two measurements per plant) were recorded. (veronica et al. 2012) 光合参数分钟尺度动态测量实验小结光合参数分钟尺度动态测量实验小结 v 实验仪器：实验仪器：li-6400（xt）6400-40 v 采样：生长采样：生长3周的叶片周的叶片 v 时间：时间：120mins v 重复数：略重复数：略 v 光：适应光：适应10分钟分钟 v 温度：略温度：略 v 二氧化碳浓度：略二氧化碳浓

34、度：略 v 湿度湿度:略略 荧光诱导曲线荧光诱导曲线 (s. devacht et al. 2011) mt = 16 ml = 100 mol m2 s1 mt = 2 ml = 100 mol m2 s1 mt = 16 ml = 800 mol m2 s1 mt = 2 ml = 800 mol m2 s1 荧光诱导曲线荧光诱导曲线 (s. devacht et al. 2011) 荧光诱导曲线解释的问题荧光诱导曲线解释的问题 v the quenching of the fluorescence signal during light induction depends on the

35、generation of npq (lambrev et al. 2007). v our results also demonstrate that the activation of npq is not only a light-dependent process (since the function of npq is to dissipate excess light energy), it is also strongly affected by temperature. v the combination of light and temperature stress aff

36、ected the psii efficiency more than each condition on its own. (s. devacht et al. 2011) 荧光诱导曲线实验方法荧光诱导曲线实验方法 v the mt was maintained throughout the whole procedure. a chl fluorescence imaging system ( cfimager, technologica, uk) was used. v the measurement procedure lasted 2.5 h (fig. 1). in a first

37、 step, the plants were dark-adapted for 30 min. (s. devacht et al. 2011) 荧光诱导曲线实验方法荧光诱导曲线实验方法 vfo was measured using a measurement light level of 0.520.85 mol m2s1. fm was measured with a saturation pulse of 4,947 mol m2s1. after 20 s, the plants were exposed to actinic light at the corresponding ml

38、 for 1 h. vduring this light period a saturation pulse of 4,947 mol m2s1was given for 800 ms every 2 min for the quenching analysis. 荧光诱导曲线实验小结荧光诱导曲线实验小结 v 实验仪器：实验仪器：cf-imager v 样品：样品：lue v 时间：暗适应时间：暗适应30分钟分钟 v 重复数：略重复数：略 v 光：暗适应；测量光：暗适应；测量fo和和fm，之后，之后20s打开打开ml v 温度：控制温度：控制 v 二氧化碳浓度：控制二氧化碳浓度：控制 v 湿度

39、湿度:略略 暗驰豫动力学暗驰豫动力学 p. lootens et al.,2011 暗驰豫动力学暗驰豫动力学 p. lootens et al.,2011 暗驰豫动力学暗驰豫动力学 p. lootens et al.,2011 暗驰豫动力学暗驰豫动力学 p. lootens et al.,2011 暗驰豫动力学解释的问题暗驰豫动力学解释的问题 vin this paper we discuss the effects of growth temperature (gt), measurement temperature (mt), and measuring light intensity (

40、ml) on the relaxation of the kautsky curve. vthe three components of the nonphotochemical quenching process (npqe, npqt, and npqi) were determined. （p. lootens et al.,2011） 暗驰豫动力学解释的问题暗驰豫动力学解释的问题 vnpqe was not affected by gt but was significantly affected by mt and ml. npqt and npqi were affected by

41、 all factors and their interactions. van acclimation effect for plants grown at low gt was detected. acclimation resulted in lower npqt and npqi values. （p. lootens et al.,2011） 暗驰豫动力学解释的问题暗驰豫动力学解释的问题 vthe halftime of the inhibition depending on npq (npqi) was not affected by any of the factors inve

42、stigated. vbased on the data generated, we conclude that npqi is a valuable parameter for screening the cold sensitivity of young industrial chicory plants. （p. lootens et al.,2011） 暗驰豫动力学实验方法暗驰豫动力学实验方法 vduring this dark period a saturation pulse was given twice after 2.5 min and every five minutes

43、thereafter during 1 h to determine the different components of the relaxation process (npqe, npqt, and npqi). 暗驰豫动力学实验方法暗驰豫动力学实验方法 vthe first two pulses after 2.5 min are important to determine npqe, the “fast” component of the relaxation process (horton and hague 1988, mller et al. 2001, bruce et a

44、l. 2004). 暗驰豫动力学实验方法暗驰豫动力学实验方法 vsubsequent pulses were applied with longer intervals, i.e., every 5 min, because the procedure can have an influence on the relaxation of npq (walters and horton 1991). the application of a saturation pulse every 5 min allowed us to maintain the influence of the pulse

45、s on the relaxation kinetics as low as possible, without any loss of information about the dark recovery. 暗驰豫动力学实验方法暗驰豫动力学实验方法 vwithin each plate 14 plants were selected at random. this corresponds to the maximum number of traces that the used chl fluorescence imaging system can record simultaneousl

46、y. chl fluorescence measurements were done on the adaxial side of the matures cotyledons. the chl signals for each of the 14 plants result from the average values of all the pixels of that individual plant. 暗驰豫动力学实验小结暗驰豫动力学实验小结 v 实验仪器：实验仪器：cf-imager v 样品：略样品：略 v 时间：暗适应时间：暗适应2.5分钟后打两次饱和闪光；之后每分钟后打两次饱和

47、闪光；之后每5分钟打分钟打1次饱次饱 和闪光，持续和闪光，持续1个小时。个小时。 v 重复数：重复数：n=14 v 光：饱和闪光光：饱和闪光 v 温度：控制温度：控制 v 二氧化碳浓度：略二氧化碳浓度：略 v 湿度湿度:略略 5 秒尺度动态变化秒尺度动态变化 v5.1 荧光参数直观成像荧光参数直观成像 v5.2 fluoresence transient 荧光参数直观成像荧光参数直观成像 (charles p. chen et al. 2011) 荧光参数直观成像解释的问题荧光参数直观成像解释的问题 vboth exposure regimes lowered leaf photosynthe

48、tic co2 uptake about 40% and psii efficiency ( fq/fm) by 20% compared with controls, but this decrease was far more spatially heterogeneous in the acute treatment. (charles p. chen et al. 2011) 荧光参数直观成像解释的问题荧光参数直观成像解释的问题 vdecline in fq/fm in the acute treatment resulted equally from decreases in the

49、 maximum efficiency of psii (fv/fm) and the proportion of open psii centres (fq/fv), but in the chronic treatment decline in fq/fm resulted only from decrease in fq/fv. (charles p. chen et al. 2011) 荧光参数直观成像解释的问题荧光参数直观成像解释的问题 vfindings suggest that acute and chronico3 exposures do not induce identic

50、al mechanisms of o3 damage within the leaf, and using one fumigation method alone is not sufficient for understanding the full range of mechanisms of o3 damage to photosynthetic production in the field. (charles p. chen et al. 2011) 荧光参数直观成像实验方法荧光参数直观成像实验方法 vattached third trifoliates were enclosed in a custom-built temperature, humidity and ambient co2-controlled gas exchange cuvette with a clear ir transmitting window (tedlar polyvinyl fluoride film, dupon