Elevated CO 2 Effects on Lichen Frequencies and Diversity Distributions in Free-Air CO 2 Enrichment ( FACE ) Station

Carbon dioxides (CO2) concentration has become much higher every year and this has already become the global issue. Lichen is a symbiotic organism that is best known as the air pollution indicator. Based on its frequency and diversity distribution, the level of pollution that has been made can be determined. This research was conducted in UKM Fernarium Free Air Carbon Dioxides Enrichment Station (FACE Station) with lichen frequencies and diversities observed and analyzed statistically and graphically. Seven species of lichens were found at the sampling locations. Findings showed that there was significant relationship in lichen frequency and diversity distribution under elevated CO2 inside FACE Station with its surrounding.


Introduction
Lichens, the product of a symbiotic relationship between a fungus (mycobiont) and at least one photosynthetic algae or cyanobacterium (photobiont), are widely used as bio-indicators of air pollution and environmental change, both at cellular and the population or community level [1].Due to the unique biology and their sensitivity to pollutants, lichens respond to variations in atmospheric composition and may be a useful measure for monitoring environmental quality.Lichens are responding to global warming and some long-term monitoring programmes have proved their sensitivity, mainly through a community approach, while only few studies investigated ecophysiological responses to climatic change [2].
In Malaysia, a limited number of studies have used lichens as indicators of air pollution, especially to evaluate lichens response on elevation of CO 2 levels [3].The negative effects of warming on the carbon balance of lichens may be at least partly counteracted by increases in atmospheric CO 2 levels.In lichens, the inability to regulate water loss limits the possible responses to CO 2 as compared to those of vascular plants, although the trade-offs related to, for instance, nitrogen allocation occur here just as much.This could also be the case for those epiphytes that grow closely attached to canopy soil or directly on living branches.For such species, increased respiration of the substrate due to higher temperatures may provide more extra CO 2 than the increased atmospheric levels.The objectives of this research were to compare the lichen diversity inside and outside Free-Air CO 2 Enrichments Station (FACE) and to analyze the distribution of identified lichen species [4] [5].

Study Area
Research was conducted in the FACE Station located in UKM Fernarium, The National University of Malaysia (02˚55'20.3"N101˚46'59.4"E).FACE Station in UKM Fernarium was programmed automatically to release CO 2 into the surroundings at fluctuated concentration.A computer-control system uses the wind speed and CO 2 concentration information to adjust the CO 2 flow rates to maintain the desired CO 2 concentration at the centre of the FACE ring.The system uses the wind direction information to turn on only those pipes upwind of the plots, so that CO 2 -enriched air flows across the plots, no matter which way the wind blows.When wind speeds are low (<0.4 m/s) and direction is difficult to detect, the CO 2 -enriched air is released from every other vertical pipe around the ring.The CO 2 flow rate is updated every second, and the choice of which vertical pipes to release from is updated every 4 seconds.Three different distances (20 m, 50 m and 80 m approximately) were selected to depict the effects of CO 2 on lichen distribution (Figure 1).

Lichen Identification
Lichen samples were collected in FACE Station and from its 80 m perimeters.All epiphytic lichen specimens were identified using a stereomicroscope and chemical spot tests, a UV lamp, and TLC following the standard checklists of Sipman in Singapore [6].

Lichen Diversity Distribution
Lichen diversity frequencies were determined using quadrate (20 cm × 50 cm).For each sampling points, 20 trees were selected randomly to evaluate lichen species distribution frequency.Lichen was sampled by each wind directions at approximately 3 meters height.Only trees with circumference around 60 -120 cm selected to conduct the sampling.Sampling was conducted in three different distances (20 m, 50 m & 80 m) [7].

Data Analysis
Every data recorded was analyzed using statistical analysis Shannon's Diversity Index and Evenness to understand the diversity's rate of lichen species at FACE Station and its surrounding.Lichen species frequency density also analyzed using graphical analysis ArcGIS 10.3 to evaluate the distribution of lichen species based on its frequency [7] [8].

Lichen Species in FACE Station
Seven species of lichen were identified and divided into three genera.All of them belong to crustose groups: Cryptothecia striata, Cryptothecia scripta, Cryptothecia granularis Cryptothecia subtecta, Porina tetracea, Myriotrema minutum and one unknown species labelled as crustose 1.All lichen been found were identified and matched with Sipman (2009) publication from Singapore.All lichen's picture that has been found in FACE station were shown in Figure 2.

Lichen Frequency Distribution
From 20 trees from each sampling locations, Table 1 shows the lichen species been found, the frequencies for each species from each location, the species diversity index (H) and the evenness value (E). Figure 3 shows the frequencies distribution for each of lichen species for all sampling locations graphically.
Based on Figure 3, the highest frequencies for all species is Cryptothecia striata with 35% at FACE Station, 21% at 50 m from FACE Station and 80 m from FACE Station.Followed by Cryptothecia granularis, Cryptothecia scripta, Porina tetracea, Myriotrema minutum, Cryptothecia subtecta and the lowest is Crustose 1 with 6.5% frequency and only can be found at 80 m from FACE Station.It shows that there are significant differences in the frequencies of lichen species in term of CO 2 effects.Increase in CO 2 concentration (inside FACE Station) unequivocally also increase the rate of photosynthesis occurred inside lichen's photobiont.In general, photosynthesis process needs CO 2 and water in order for the process run in optimal condition.That's the reason why frequencies of lichen species that exist inside FACE Station were much higher compare to 50 m from FACE Station and 80 m from FACE Station.
In term of diversity, different CO 2 concentration affected lichen diversity substantially.Lichen diversity increased gradually when receding the FACE Station.According to species diversity index (H) in Table 1, the value of the index increased from FACE Station is 1.35, 50 m from FACE Station 1.53 and 80 m from FACE Station is 1.89.Increase in CO 2 concentration affected several of lichen's growth such as Cryptothecia scripta, Cryptothecia subtecta and Crustose 1.These three species only existed outside the FACE Station.The difference in lichens distribution shows that lichens get affected with the elevation of CO 2 .The elevation of CO 2 concentration acidified the bark of tree which is important for epiphytic lichen attachment.The rhizine of lichen only recognize particular pH of substrate.Changes in pH bring no growth for that particular of lichens.

Conclusion and Recommendation
CO 2 concentration has various effects on lichen distribution in terms of frequency and diversity.Lichen frequencies

Figure 2 .
Figure 2. Lichens from FACE station, 20 m and 50 m.areproportional with CO 2 concentration and lichen diversity is inversely proportional with CO 2 concentration as showed in this research.For further research, we should look up into lichen physiology, for example, chlorophyll a and b which are important in photosynthesis.This may give a brand new perspective and precise observation into how elevation of CO 2 affects on lichen physiologically.

Figure 3 .
Figure 3. Frequencies distributions for each lichen species.

Table 1 .
Lichen frequency distribution in FACE Station.