EN 16789:2016

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.Zunanji zrak - Biomonitoring z višjimi rastlinami - Metoda standardizirane izpostavljenosti tobakaAußenluft - Biomonitoring mit Höheren Pflanzen - Verfahren der standardisierten Tabak-ExpositionAir ambiant - Biosurveillance à l'aide de Plantes Majeures - Méthode de l'exposition de tabac standardiséeAmbient air - Biomonitoring with Higher Plants - Method of the standardised tobacco exposure13.040.20Kakovost okoljskega zrakaAmbient atmospheresICS:Ta slovenski standard je istoveten z:EN 16789:2016SIST EN 16789:2017en,fr,de01-julij-2017SIST EN 16789:2017SLOVENSKI
STANDARD
EUROPEAN STANDARD NORME EUROPÉENNE EUROPÄISCHE NORM
EN 16789
August
t r s x ICS
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s uä r v rä t r English Version
Ambient air æ Biomonitoring with Higher Plants æ Method of the standardized tobacco exposure Air ambiant æ Biosurveillance à l 5aide de plantes supßrieures æ Mßthode de l 5exposition normalisße du tabac
Außenluft æ Biomonitoring mit Höheren Pflanzen æ Verfahren der standardisierten TabakæExposition This European Standard was approved by CEN on
s z June
t r s xä
egulations which stipulate the conditions for giving this European Standard the status of a national standard without any alterationä Upætoædate lists and bibliographical references concerning such national standards may be obtained on application to the CENæCENELEC Management Centre or to any CEN memberä
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t r s x CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Membersä Refä Noä EN
s x y z {ã t r s x ESIST EN 16789:2017

Reference plates and photographs for evaluating the percentage of necrosis on leaf surfaces . 24 Annex B (informative)
Documentation . 28 B.1 General . 28 B.2 Example of the information that shall be recorded at a given exposure location . 28 B.3 Example of the information that shall be recorded for a tobacco plant at a given assessment date . 30 Bibliography . 31 SIST EN 16789:2017

Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. SIST EN 16789:2017

« x r r mg/l. NOTE The NPK-content of commercial potting soils is frequently given as weight per litre of the product. As such, further fertilisation during cultivation and exposure of the bioindicator plants is not necessary. The substrate should have a pH between 5,5 and 6,5. Before putting the soil into the plant pots, it should be moistened if necessary. 4.1.3 Water For watering the plants drinking water quality (Council Directive 98/83/EC on the quality of water intended for human consumption [63]) is sufficient. If the values given there cannot be complied with, deionised water shall be used. 4.1.4 Exposure device The exposure of the bioindicators takes place in commercially available square plastic plant pots with the dimensions 13 cm × 13 cm (top rim) and a height of 13 cm (volume: ca. 1,25 l to 1,5 l; see Figure 1) or in round pots with comparable soil volume. Four holes are drilled into the base of the pots (if not already present in the purchased pots), through which two moistened glass fibre wicks (diameter: 5 mm to 6 mm, length: 50 cm to 70 cm) or other suitable suction wicks are inserted. The wicks serve as a continuous water supply during cultivation and exposure. At least 7 cm of the wicks should reach into the substrate. The length of loose ends should be chosen in such a way that both ends reach the bottom of the water storage container. As water storage container, a Euro standard stackable plastic container (60 cm × 40 cm × 12 cm) is used, into which an overflow is drilled approximately 2 cm below the upper edge. A white polystyrene block (60 cm × 40 cm × 11 cm) with at least two recesses (11,5 cm × 11,5 cm) into which the plant pots are put is placed onto this tub. In this way, mutual obstruction/shading of the growing plants is avoided. Suitable shaping of its lower edge prevents the block slipping off the tub (Figure 1). For the plants, wooden or bamboo sticks are used as support to prevent wind damage. SIST EN 16789:2017

Key 1: leaf with necrosis 2: polystyrene block 3: water reservoir 4: suction wick Figure 1 — Example of a device for the exposure of tobacco plants 4.1.5 Exposure rack The exposure rack consists of a solid frame construction (Figure 2). The tobacco plants are exposed at a height of 70 cm to 110 cm from ground level to the soil surface in the pots. During the exposure water is supplied by the wicks, which hang from the plant pots into the water reservoir. A filling quantity of 20 l enables two weeks of maintenance-free exposure. The exposure rack is covered with green shading fabric (shading rate 50 %) at the top and at three sides (east, south, west). It is open toward the north. The shaded plants react more sensitively to ground-level ozone than those under direct sunlight as the stomata of the leaves – being the dominating uptake path for ozone – remain open for longer periods of time. One can therefore expect a higher level of leaf injury in shaded plants. SIST EN 16789:2017

Key 1: metal frame construction, consisting of four frame elements (2 pieces 180 cm × 90 cm, 2 pieces 180 cm × 60 cm) 2: shading fabric at three sides (east, south, west) as well as at the top 3: stackable plastic containers (60 cm × 40 cm × 12 cm) as water storage containers 4: example of a polystyrene block (60 cm × 40 cm × 11 cm) with three complete pot-shaped recesses (11,5 cm × 11,5 cm) and recessed edge to prevent slipping 5: plastic pot (13 cm × 13 cm) with suction wicks Figure 2 — Exposure rack [SOURCE: [64]; modified] 4.2 Cultivation of plants The aim of the cultivation is a healthy, vigorous plant. The tobacco plants are cultivated in an environment which minimizes ambient ozone concentration (e.g. greenhouse, open-top chamber, phytotron, if possible supplied with charcoal-filtered air). Efforts should be made to ensure as constant temperature as possible during cultivation. At high temperatures the seedlings should be watered from top down using a sprinkler, in order to avoid overheating; at night the temperature should not drop below 10 °C. The aim is to produce plants at a comparable stage of development, and thus similar sensitivity, for all exposure periods. The tobacco is sown in trays with well-moistened standardized soil (see 4.1.2). The top layer consists of sieved soil, which is smoothed to a flat surface. The seeds are applied evenly and spread very thinly (1 seed/cm2 to 5 seeds/cm2). Since tobacco plants require light for germination, the seeds are not covered with soil. The seed trays are maintained at temperatures between 18 °C and 28 °C, in order to ensure a safe and rapid germination within five to 16 days. If higher temperatures occur, the trays should be kept in a shady place to avoid heat stress. During this time, the surface of the soil shall always be kept moist (e.g. under a transparent cover on the tray). Unused seeds can be stored in a refrigerator (4 °C to 6 °C) for future use (see 4.1.1). After the initial growth, the seedlings (see Figure 3a) should be transferred to a place with ample light. SIST EN 16789:2017

Figure 3a — Seedlings Figure 3b — Two leaf-stage The following cultivation procedure is recommended: Two to three weeks after sowing, the seedling reaches the two leaf-stage (two successive leaves after the two cotyledons; the latter do not count as leaves; see Figure 3b). At this stage, four small bunches of two to six seedlings each are taken from the sowing trays and transferred into the exposure pots (Figure 4). The pots should be kept in a shady place for five to seven days to allow proper root growth without heat and drought stress. Thereafter, a sunnier place is favourable for further growth.
Figure 4 — Tobacco seedlings, immediately after potting The plants are watered carefully the first time using a thin spout to avoid damage to the seedlings. The pots are put into a water storage container without the polystyrene block and set up in a protected, relatively shaded place. The bottom of each container is covered with some water, so that the soil in the pots always remains moist by the use of the suction wicks. In the next two to three days it is checked whether at least one plant from each of the four bunches has established (areas where no plant has established satisfactorily should be replanted). One week after the replanting at the latest, four seedlings out of the bunches are selected in such a way that in each pot remain four plants of different sizes (Figure 5). SIST EN 16789:2017

Figure 5 — Tobacco plants after selection to four plants/pot Five to six weeks after sowing, the plants will have reached a stage at which they begin to compete with each other. They are therefore thinned to one plant per pot (Figure 6). In doing so, either the largest, middle or smallest plant of each pot is left for exposure – depending on the general developmental stage (in order to get a set of plants as homogeneous in size and development as possible).
Figure 6 — Single tobacco plant Once the plants have reached the stage depicted in Figure 7, they are exposed at the monitoring location (soil should be well moistened beforehand). Before exposure at the latest, each plant shall be loosely tied to a wooden or bamboo stick (of at least 40 cm). SIST EN 16789:2017

Key left: leaf pattern of a tobacco plant; leaves 2 to 8 right: leaf pattern of a tobacco plant; leaves 1 to 8; leaves coloured grey = those used for visual injury assessment; a = ring added to mark the lower bound above which leaves are assessed Figure 7 — Tobacco plants at the exposure stage Leaf pattern of a plant ready for exposure (the leaf length refers to the blade only, not including the petiole): — The two cotyledons have already fallen off and are not counted. — Leaf 1 has reached a maximum size of approximately 2 cm–3 cm in diameter; it remains round. — Then follows a likewise round leaf of approximately 3 cm–5 cm maximum diameter (No. 2). — The next leaf (No. 3) is oval, bluntly pointed and reaches a maximum of hand size when fully grown. — The next leaf (No. 4) is the first leaf that features the typical tobacco leaf shape (pointed oval) and is substantially larger than leaf No. 3. Above this leaf a reference ring or a similar indicator is fitted at the beginning of the exposure. This indicator marks the lower bound above which visual injury assessment begins (see Clause 5). The exposure stage is considered to be reached when leaf 8, the fourth leaf above the reference ring, has a length of at least 10 cm, while leaf 9 is still shorter than 10 cm in total length. This stage is reached seven to 10 weeks after sowing depending on the local climatic conditions. A summary of the cultivation is given in Table 1. SIST EN 16789:2017

Exposure location 1 Exposure location 2 Exposure location 3 Leaf number 5 6 7 5 6 7 5 6 7 Plant 1 10 10 5 40 10 5 50 20 0 Plant 2 5 5 5 20 20 5 X 50 10 Plant 3 20 30 5 40 20 5 X X 30 Plant 4 10 10 5 50 20 10 40 50 20 Plant 5 20 10 10 X 30 10 60 40 20 6.2.3 Missing value completion Table 2 is expanded by adding a line below that contains the mean values for each location and leaf position that have not lost any reference leaves. In the example data set, this mean is based on five plants for Location 1, four plants for Location 2 and three plants for Location 3 (Table 3). If the data set contains no missing values, the data processing may proceed as shown in Table 5. SIST EN 16789:2017

Exposure location 1 Exposure location 2 Exposure location 3 Leaf number 5 6 7 5 6 7 5 6 7 Plant 1 10 10 5 40 10 5 50 20 0 Plant 2 5 5 5 20 20 5 X 50 10 Plant 3 20 30 5 40 20 5 X X 30 Plant 4 10 10 5 50 20 10 40 50 20 Plant 5 20 10 10 X 30 10 60 40 20 Means of plants without missing leaves 13,0 (65/5) 13,0 (6
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