Force-A Multiplex Use

display these 4 usual signal/indices on the touch screen : Check Multiplex every day you measure with the blue (and red for Mx 330) standard : use the sheet ‘Multiplex checking’. You must avoid saturation in single signals. FRF_R, FRF_G, FRF_B, FRF_UV must not : - exceed 4300 - be under 20 You have to adapt the « measurement range » to: 1/ diameter of the aperture of Multiplex. 2/ level of chlorophyll of the plant you are measuring 3/ in field or in greenhouse measurements For more explanations, see MxUserGuide_Firmware.pptx. Slide 6 : Cfg screen. Mx 330: use black glove to measure

For both measurement mode (one shot and continuous), 3 principles to respect: - measure leaves with the same physiological age - try to be as close as possible to the leaves - try to cover as much as possible the aperture of the Mx with leaves

To be sure of the quality of the signals, use the measurement area well adapted to the plant you measure : 4 / 6 or 8 cm aperture diameter. NO OK You must always keep the same distance from the fruit. The good distance is : the fruit against the Multiplex. OK NO Be careful with the pedicel of a fruit. It is rich in chlorophyll, it influences the reading.

except under rain. You must not measure with the Multiplex turned towards the sun. NO NO NO Avoid fluorescent light if you measure in the laboratory.

- Wash the measurement area and the plastic protection upon optics with alcohol or water as soon as these parts are dirty. - Look at FRF_R during measurement. Be sure it is < 4300 and > 20. - Compile measurements in the air and on the blue standard every day in a dedicated file. - Try to do the same plot at the same time every day, in case of measurement of kinetics. - User effect is possible. Try to always dedicate one Mx to the same user. - Don’t exceed 100 files on the SD card. Transfer and delete files regularly . - Use the 512 Mo SD card provided with the Multiplex.

Measurement can be performed by pressing one of the two trigger buttons on the handle.

screen »: to come back on previous ‘p’ ‘s’ or ‘file name’ Access to « Cfg screen »: to settle different settings Access to « Del screen »: to delete last reading => 4 levels to classify your measurements .csv file column: population sample reading .csv file name Battery level Time One signal with its value. You can display all the signals saved by Multiplex, if you press successively here. When GPS is on, this symbol is displayed. ‘population’ / ‘sample’ / ‘file name’ are long press buttons (2 sec) : you can increment the rank by pressing the respective buttons.

- YY : year - MM : month - DD : day - XX : two letters which correspond to the file name (bottom-right edge of the main screen) A new file is created each time you : - switch on the Multiplex, - press the file name button, When a new file is created : - there are incrementation of the 2nd letter of the file name - ‘p’ and ‘s’ rank are back to 1 - ‘reading’ rank is back to 0. The reading rank in each file is not limited. Save regularly the files on a computer and delete them from the SD card. All datas are saved on the sd card, into a .csv file.

have to connect it on Mx if you want to use the GPS mode. External GPS : if you want to use your own GPS Another screen allow you to set the Baud rate (4800, 9600,…) On / Off : Display if the GPS is on or off. Mode 1 : To switch on GPS, press this button and wait few seconds (only outdoors). For each measurement, GPS datas are saved on .csv file. Mode 2 : it appears when ‘measurement mode’ is ‘contin.’ In this mode, only 2 GPS coordonnates are saved, at the beginning and the end of the continuous measurement Acquisition speed of ‘continuous’ mode : Mode 1 = 2 Hz / Mode 2 = 20 Hz. Mode 2 If the GPS is connected, the date and time are updated.

main --> One shot One shot mode (250 flashes per reading) Continuous mode (acquisition speed : 20 Hz) Datas saved on the sd card Datas saved on a PC with a USB cable Continuous mode : only raw single signals are saved in the file (to allow the acquisition speed = 20 Hz). Indices need to be calculated separately. You can use the file MxContinuousMode_Formula.xlsx to get the formulas directly.

(Cfg 4 can appear for a few customers : usually Cfg 2 < Cfg 4 < Cfg 3) The aim of these 3 ‘measurement range’ is to get enough fluorescence, but not too much. FRF_R, FRF_G, FRF_B, FRF_UV (when UV LEDs present) must not exceed 4300 and be under 20. You have to adapt the ‘measurement range’ to: 1/ diameter of the aperture of Multiplex. 2/ level of chlorophyll of the plant you are measuring 3/ in field or in greenhouse measurements 4/ motionless or in movement measurement Usually, for the aperture diameter 6 / 8 cm, motionless : Cfg 1 : plant in greenhouse Cfg 2 : leaves, fruit unripe outdoors. Cfg 3 : mature fruit with low level of chlorophyll Usually, for the aperture diameter 4 cm, motionless : Cfg 2 : plant in greenhouse Cfg 3 : leaves, fruits

Display one or four signals on the touch screen Adjust time with your local time (use this button if you don’t use the GPS) One-signal-displayed mode speeds up a little bit measurements compared to the four- signals-displayed mode.

last measure ? Yes To delete the last measurement saved on the sd card.

You just have to adjust with ‘+’ or ‘–’ the level of classification you want to pursue. Alt screen is very useful if you have to switch off the device (lunch time,…) during a measurement session. Measurement rank can not be changed.

17 1 2 3 4 Leave Face Plane Tree Ad Plane Tree Ab Hazel Ad Hazel Ab SFR_R FLAV ANTH_RG 2,11 0,40 -0,04 1,13 -0,05 -0,01 2,34 0,46 -0,01 0,97 -0,17 0,11 1 2 3 4 The Multiplex measurement area allow to integrate the leaf heterogeneity. SFR_R: Chlorophyll index. Values are different between adaxial and abaxial side because chlorophyll content is higher in adaxial side (palisade parenchyma) than in abaxial side (spongy parenchyma) FLAV: Flavonols index. Values are higher in adaxial side compared to abaxial side (UV light induce flavonols). Like here, FLAV can be < 0. FLAV = log (FRF_R / FRF_UV), when there is few flavonol in the leaf, FRF_UV > FRF_R. ANTH_RG: Anthocyanins index (for red leaves, like in picture 4). In the absence of anthocyanins, ANTH_RG is inversely correlated to chlorophyll content. Index range values : SFR_R : 0 to 3. FLAV : - 0,5 to 2,5. ANTH_RG : -0,2 to 1,5. Values are relative values. They can’t be completely compared between species. Multiplex always displays a value, whatever the support (air, table,…), and even if there is no chlorophyll, polyphenols,… Multiplex can’t measure on liquid (the way to measure need to respect the scheme : epiderm / mesophyll).

We can see differences between : - Cultivars - Readings on sun-exposed or shaded side Multiplex readings on fruits. SFR_R, ANTH and FLAV are corrected by the value of blue standard foil reference.

19 Pisa University. 2010. 6 treatments : 0 / 50 / 100 / 150 / 200 / 250 kg N / Ha. 28th of june. 4 replicates (1 * 1 m) per treatment. Mx measurement. 12th of July. Leaf N content. Agati et al. EJA . 2013. NBI® NBI® = CHL / FLAV