Aerobiologia DOI 10.

1007/s10453-011-9197-z

ORIGINAL PAPER

Mapping airborne pollen of papaya (Carica papaya L.) and its distribution related to land use using GIS and remote sensing
Apichart Sritakae Patchara Praseartkul Wasinee Cheunban Poonsak Miphokasap Apisit Eiumnoh Parichart Burns Namthip Phironrit Bencharong Phuangrat Panit Kitsubun Asawin Meechai

Received: 25 January 2010 / Accepted: 12 January 2011 Springer Science+Business Media B.V. 2011

Abstract Papaya is an economically important plant in Thailand for domestic consumption and export. However, papaya is extremely susceptible to disease caused by the papaya ring spot virus. Although transgenic papaya has been developed, commercial cultivation of transgenic plants in Thailand is still illegal. One concern is cross-pollination to conventional varieties. In this study, windborne-pollen dispersion of papaya (Carica papaya L.) was investigated using geographic information systems (GIS) and remotely sensed data. Pollen traps were placed around a papaya plot in eight geographic directions, with radiuses varying from 5 to 900 m from the plot. Pollen counts were made for 12 different dates, and data were input into a GIS

database. The distribution of pollen and its relation to land use were analyzed using land use data obtained from Quickbird imagery acquired during 2007. Comparative analyses of pollen dispersal, wind direction, and speed were made using data collected from a micro-climatic station set up at a papaya plot. The furthest distance from the plot that pollen was found was at 0.9 km, a distance at which only 1 pollen grain was found. The number of pollen grains carried by wind decreased as distance increased. The direction of dispersal was not in accordance with wind direction data. Most pollen grains were found in agricultural areas and bare land. The total number of pollen grains found in exposed areas was considerably higher than the total found in areas sheltered by dense tree lines.

A. Sritakae (&) P. Praseartkul W. Cheunban P. Miphokasap A. Eiumnoh Geoinformatics Laboratory, BIOTEC Central Research Unit, National Centre for Genetic Engineering and Biotechnology, Thailand Science Park, Pathumthani, Thailand e-mail: apichart@hotmail.co.uk P. Burns N. Phironrit Plant Research Laboratory, Agricultural Biotechnology Research Unit, National Centre for Genetic Engineering and Biotechnology, Thailand Science Park, Pathumthani, Thailand P. Burns B. Phuangrat Center for Agricultural Biotechnology, Kasetsart University, Kamphaeng Saen, Nakhon Pathom, Thailand

P. Burns B. Phuangrat Center for Excellence on Agricultural Biotechnology (AG-BIO/PERDO-CHE), Bangkok, Thailand A. Meechai King Mongkuts University of Technology Thonburi, Bangmod, Toongkru, Bangkok, Thailand P. Kitsubun A. Meechai Biochemical Engineering and Pilot Plant Research and Development Unit, National Centre for Genetic Engineering and Biotechnology, King Mongkuts University of Technology, Thonburi, Bangmod, Toongkru, Bangkok, Thailand

123

Aerobiologia

Keywords Papaya Pollen Spatial dispersal GIS Remote sensing

1 Introduction Papaya is a very popular fruit in Thailand. It is used in various traditional recipes, such as spicy green papaya salad. In addition to domestic consumption, Thailand exports papaya to 21 countries worldwide (Department of Customs 2008). Papaya is polygamous species consisting of three sex forms: male, female, and hermaphrodite (Eustice et al. 2008). In general, the hermaphrodite form is most preferable for commercial uses because of its desirable fruit shape (Yu et al. 2008). Papaya hermaphrodites bear elongated owers, although high temperatures can result in the reduction of ovaries and cause the ower to function as male. This is known as reduced elongata (Nakasone and Paull 1998). Papaya fruiting occurs via cross-pollination, selfpollination, or parthenocarpy (Teixeira da Silva et al. 2007). Pollen production and pollination occur year round; however, pollen viability and pollination efciency decline during the summer (as observed by the number of setting seeds). Anther of reduced elongata and elongata owers are dehisced 2 days and 1 day before ower blooming, respectively. In addition, stigmas are receptive 3 days before blooming and have maximum receptivity to the fth day of anthesis (OECD 2005). Plants start owering at a height of 1.2 m. Flowers bloom both in the morning (OECD 2005) and in the evening (Pares et al. 2002; Sippel et al. 1989). Papaya is extremely susceptible to the papaya ring spot virus (PRSV). In Thailand, the PRSV was discovered in 1975 and affects all growing areas, especially the northeast. In 1996, an effort to solve this problem was made by Kasetsart University and the National Center for Genetic Engineering and Biotechnology (BIOTEC) through the development of a transgenic papaya that is resistant to PRSV. However, the release of transgenic papaya to the environment is illegal in Thailand because of safety concerns for the public. In particular, there are concerns about potential contamination by cross-pollination to non-GM crops. A recent issue of interest is pollen dispersion and pollen concentration that may cause contamination from one species to another. For instance, seed producers concerns about the purity of their seed products recently reemerged with the development of transgenic plants

(Treu and Emberlin 2000). Numerous studies investigated and modeled the airborne-pollen dispersion of crops such as wheat, corn, potatoes, sugar beets, soybean, and rape seed for oil (Abud et al. 2007; Bannert and Stamp 2007; Bateman 1947; Ivanovska et al. 2009; Loos et al. 2003; Paterniani and Short 1974; Raynor et al. 1972; Schefer et al. 1993; Scott 1970; Skogsmyr 1994; Timmons et al. 1995; Tynan et al. 1990; Wagner and Allard 1991). The dispersal distances found in these studies are varied and depend on several relevant factors, including climatic conditions, topography, and land use. However, there are no published data on papaya pollen dispersion. This is probably because papaya is less consumed compared with other major food crops. The objectives of this research are to investigate the dispersion of papaya pollen and its distribution in relation to land use and wind speed and direction using geographic information systems (GIS) and remote sensing as analysis tools.

2 Materials and methods 2.1 Seedling preparation and experimental plot establishment The study was carried out in 20072008 on the Kamphaengsaen campus of Kasetsart University. Due

Fig. 1 Layout of papaya seedlings planted in the experimental plot. Hermaphrodites were planted at the center of the plot, with female plants surrounding them

123

Aerobiologia

to the fact that releasing transgenic plants to the environment is illegal in Thailand, non-transgenic papaya (Carica papaya L.) was used for this experiment. However, morphological characteristics (weight, size, surface and form) of transgenic pollen and non-transgenic pollen of this variety are reportedly comparable (BIOTEC unpubl. data). Seedlings were prepared in a greenhouse. Papaya seeds were planted in a medium consisting of coconut ber, soil, and cattle manure in proportions of 1:1:1, respectively. Four-week-old seedlings were later transplanted with 2 9 2 m spacing on a 30 9 32 m experimental plot. Hermaphrodite plants were planted at the center of the plot as a pollen source, and female plants were planted around them

(illustrated in Fig. 1). The 46-0-0 fertilizer was applied at an early stage until plants started fruiting; 16-16-16 fertilizer was then applied twice monthly. 2.2 Pollen and climatic data collection To measure papaya pollen dispersion, a pollen trap was designed for this study. A gypsum plate was rmly nailed at its center to a wooden stake. On it, a very thin double-sided adhesive paper (6.5 9 11.5 cm) was xed with a rectangular plastic board that was used as an airborne-pollen collector (Fig. 2). Pollen traps were installed around the papaya plot in eight directions (N, NE, E, SE, S, SW, W, and NW) and at various distances (5, 15, 30, 50, 80, 100, 200,

Fig. 2 The pollen trap consisted of a measurement station made from a solid gypsum board centrally nailed to the top of a wooden stake, (a) and a pollen collector made from double-

sided adhesive paper attached to a plastic board (b), which was put on top of the measurement station

123

Aerobiologia

300, 400, 500, 600, 700, 800, and 900 m; illustrated in Fig. 3). The height of the pollen measurement station from the ground to the top was set to 1.5 m, which is close to the height at which papaya plants ower. The locations of the plot and traps were input into a geographic information system (GIS) using the software ArcGIS 8.0 (ESRI 1999). Prior to measurements used for the analyses, the system was calibrated several times using the same methods as the actual data collection. This ensured that there was no contamination of papaya pollen from other sources. Moreover, the isolation of the plot with regard to other papaya plots was conrmed by intensive eld checks using maps produced from

satellite imagery. There was only 1 other older papaya plot in the vicinity; it was approximately 1 km away from the study plot and was already inactive in pollination. We took a total of 12 measurements. Each pollen measurement was taken either from 7:0016:00 h or from 17:0007:00 h in accordance with the pollination periods in the morning and evening (OECD 2005; Sippel et al. 1989; see Table 1). For each measurement, pollen collectors were placed at all stations and were left for the duration of the hours of pollination previously mentioned. Subsequently, the pollen collectors were removed from the measurement stations, and the adhesive papers were immediately xed on slides.

Fig. 3 Quickbird satellite imagery acquired in 2007 showing the study area on the Kamphaengsaen campus of Kasetsart University. A papaya plot was laid out, and pollen measurement stations were installed around the papaya plot in 8 directions at various distances

123

Aerobiologia Table 1 Dates and times of 12 pollen measurements Date June 23, 2008 June 24, 2008 July 1, 2008 July 2, 2008 July 7, 2008 July 8, 2008 July 22, 2008 July 22, 2008 July 29, 2008 July 30, 2008 August 4, 2008 December 11, 2008 Time of measurement 17:007:00 h 7:0016:00 h 7:0016:00 h 7:0016:00 h 17:007:00 h 7:0016:00 h 7:0016:00 h 17:007:00 h 17:007:00 h 7:0016:00 h 17:007:00 h 17:007:00 h

created for each measurement using WRPLOT VIEW version 5.9 (Lake Environment 1998). 2.3 Land use data acquisition To investigate the relationship between pollen dispersion and land use, a land use map was produced from 0.6 m resolution. Quickbird multispectral data acquired March 3, 2007. ENVI 4.3 (ITT Visual Information Solutions 2006) was used to process the satellite data. Georeferencing was conducted using 32 ground control points with a second-order polynomial transformation. The result produced a satisfactory RMS (root mean squared error) of 0.001 m. Ground truth data were collected and used as training areas for a supervised classication and accuracy assessment. We then performed a supervised classication of 3 spectral bands (blue: 450520 nm; green: 520600 nm; and red: 630690 nm) using a maximum likelihood classier. The classication resulted in an acceptable land use map (overall accuracy = 77.9%; Kappa coefcient = 0.7173). There were some misclassied areas caused by similar spectral intensity of some land use classes. We improved the accuracy of the land use map by conducting intensive eld checks and correcting misclassied data. Raster land use data were then converted to vector data and overlaid with the pollen dispersion data to analyze their relationship.

Slides were marked with date, time, and position, and then they were safely kept in a slide tray for ease of transportation and protection from contamination. Slides were then stained with Safranin O, and pollen was counted under a light microscope at 209 magnication. Pollen count data were then input into a GIS database, and a pollen dispersion map was produced for all measurements to compare results with climatic data. Climatic data, wind speed, and direction were recorded every 5 min using a micro-climatic station installed at the center of the papaya plot, as illustrated in Fig. 4. To analyze the effects of wind speed and direction on pollen dispersion, wind roses were

Fig. 4 Micro-climatic station installed at the center of the papaya plot. The station recorded wind speed and direction every 5 min

Fig. 5 The relationship between total pollen and distance from the pollen source. Total pollen decreased as distance increased. The number labels on the red dots show the variance of the 12 measurements at each distance; these are high at short distances and rapidly decrease at longer distances

123

Aerobiologia

3 Results and discussion 3.1 Spatial patterns of pollen dispersion The results showed that pollen dispersion extended up to 900 m from the source, and the total pollen decreased with increasing distance from the papaya plot. Beyond 100 m, total pollen counts rapidly decreased. The variability of the pollen count was very high at short distances and was lower at longer
Fig. 6 The comparison between pollen dispersion and wind speed, and direction were organized according to date of measurement. The top image of each measurement is a wind rose plot representing the distribution of wind speed and direction. Color bands show the wind direction and speed from a micro-climatic station installed at the center of the papaya plot. The length of the color bands indicates the frequency of that direction. The lower image represents pollen dispersion measured. Most measurements show that pollen distribution was not in accordance with wind speed and direction

distances (see Fig. 5). Interestingly, visually comparative analyses showed that most of the pollen distribution recorded was not in accordance with wind speed and direction (Fig. 6). This was probably caused by the very low wind speed in the Nakhon Pathom province (the studied area), where very low annual wind speed (\2 m/s, dened as calm wind) prevails (Department of Alternative Energy Development and Efciency 2007). This is common in the tropics (Sharan et al. 1996), and under low wind

123

Aerobiologia Fig. 6 continued

speeds, the dispersion of airborne particles is irregular and not always predictable (Carvalho and Vilhena 2005). Consequently, wind direction and airborne dispersion under such conditions are difcult to describe (Anfossi et al. 2005). Apart from wind speed and direction, other factors, such as seasonal anomalies of pollen shedding and climatic dynamics, should be taken into consideration for future studies.

3.2 Land use and pollen dispersion The results show eight principle types of land use in the studied area. Total pollen in agricultural areas was higher than in other land use types. The second greatest amount of pollen fell onto bare land or cleared areas after harvest. Interestingly, there was considerably less pollen behind lines of dense tree

123

Aerobiologia Fig. 6 continued

stands (aligning from the north to the southwest) than in other areas (illustrated in Fig. 7). This result is similar to that found in the study by Goodwille (1999), who reported that airborne-pollen counts in exposed areas were different from those in sheltered sites. Treu and Emberlin (2000) reviewed relevant studies and reported that vegetation barriers such as dense hedges or trees depleted airborne pollens,

altered the impact of airow, and generated downwind sheltered zones.

4 Conclusions As a eld trial, this study showed that the dispersion of airborne papaya pollen under low wind speed

123

Aerobiologia Fig. 7 The distribution of pollens related to land use shows that most pollen grains were found in agricultural areas. Very little pollen was found in areas behind dense tree lines aligning from the north to the southwest

conditions was not in accordance with wind direction. Pollen grains were densely distributed within 100 m from the papaya plot and dramatically decreased beyond that range. The farthest distance where pollens were found was 900 m from the plot. Total pollen varied in different land use types; the types with the most pollen were agricultural areas, bare land, and harvested areas. Moreover, areas sheltered by dense tree lines had considerably less pollen than exposed areas. The simple methods used in this study demonstrate the basic implications of real-life scenarios relevant to cross-pollination of transgenic plants to conventional varieties.

Acknowledgments We acknowledge the Cluster and Program Management Ofce (CPMO) at the National Center for Science and Technology Development (NSTAD) for nancial support of this research. We thank the National Electronics and Computer Technology Center (NECTEC) for providing the micro-climatic station used in this study. We also would like to thank the Agriculture Biotechnology Center (CAB) at Kasetsart University for supporting eld work.

References
Abud, S., de Souza, P. I. M., Vianna, G. R., Leonardecz, E., Moreira, C. T., & Faleiro, F. G. (2007). Gene ow from transgenic to nontransgenic soybean plants in the Cerrado region of Brazil. Genetic and Molecular Research, 6, 445452.

123

Aerobiologia Anfossi, D., Olettl, D., Degrazia, G., & Goulart, A. (2005). An analysis of anemometer observations in low wind speed conditions. Bound-Lay Meteorol, 114, 179203. Bannert, M., & Stamp, P. (2007). Cross-pollination of maize at long distance. European Journal of Agronomy, 27, 4451. Bateman, A. J. (1947). Contamination in seed crops 2. Wind pollination Heredity, 1, 235246. Carvalho, J. C., & Vilhena, M. T. (2005). Pollutant dispersion simulation for low wind speed condition by the ILS method. Atmospheric Environment, 39, 62826288. Department of Alternative Energy Development and Efciency. (2007). Wind resource assessment of Thailand. Available at: http://www2.dede.go.th/dede/renew/Twm/ main.htm. Accessed Jan 2010. Department of Customs. (2008). Export statistics. Available at: http://www.customs.go.th. Accessed Jan 2010. Eustice, M., Yu, Q., Lai, C. W., Hou, S., Thimmapuram, J., & Liu, L. (2008). Development and application of microsatellite markers for genomic analysis of papaya. Tree Genetics and Genomes, 4, 333341. Goodwille, C. (1999). Wind pollination and reproductive assurance in Linanthus parviorus (Polemoniaceae), a self-incompatible annual. American Journal of Botany, 86, 948954. Ivanovska, A., Todorovski, L., Debeljak, M., & Dzeroski, S. (2009). Modelling the outcrossing between genetically modied and conventional maize with equation discovery. Ecological Modelling, 220, 10631072. Loos, C., Seppelt, R., Meier-Bethke, S., Schiemann, J., & Richter, O. (2003). Spatially explicit modeling of transgenic maize pollen dispersal and cross-pollination. Journal of Theoretical Biology, 225, 241255. Nakasone H.Y., Paull R.E.(1998).Tropical fruits. CAB International, Wallingford. OECD (Organisation for the Economic Cooperation and Development). (2005). Consensus document on the biology of papaya (Carica papaya). series on harmonisation of regulatory oversight in biotechnology No. 33. Paris: OECD Environment Directorate. Pares, J., Basso, C., & Jauregui, D. (2002). Momento de antesis, dehiscencia de anteras y receptividad estigmatica en ores de lechosa (Carica papaya L.) cv. Cartagena Amarilla. Bioagro, 14, 1724. Paterniani, E., & Short, A. C. (1974). Effective maize pollen dispersal in the eld. Euphytica; Netherlands Journal of Plant Breeding, 23, 129134. Raynor, G. S., Ogden, E. C., & Hayes, J. V. (1972). Dispersion and deposition of corn pollen from experimental sources. Agronomy Journal, 64, 420427. Schefer, J. A., Parkinson, R., & Dale, P. J. (1993). Frequency and distance of pollen dispersal from transgenic oilseed rape (Brassica napus). Transgenic Research, 2, 356364. Scott, R. K. (1970). The effect of weather on the concentration of pollen within sugar-beet crops. The Annals of Applied Biology, 66, 119127. Sharan, M., Singh, M. P., & Yadav, A. K. (1996). Mathematical model for atmospheric dispersion in low winds with eddy diffusivities as linear functions of downwind distance. Atmospheric Environment, 30, 11371145. Sippel, A. D., Claassens, N. J. F., & Holtzhausen, L. C. (1989). Floral differentiation and development in Carica papaya cultivar Sunrise Solo. Scientia Horticulturae, 40, 2333. Skogsmyr, I. (1994). Gene dispersal from transgenic potatoes to conspecics: A eld trial. Theoretical Applied Genetics, 88, 770774. Teixeira da Silva, J. A., Rashid, Z., Nhut, D. T., Sivakumar, D., Gera, A., & Souza, Jr. M. T. (2007). Papaya (Carica papaya L.) biology and biotechnology. Tree and Forestry Science and Biotechnology, 1, 4773. Timmons, A. M., OBrien, E. T., Charters, Y. M., Dubbels, S. J., & Wilkinson, M. J. (1995). Assessing the risks of wind pollination from eld of genetically modied Brassica napus ssp. oleifera. Euphytica, 85, 417423. Treu, R., & Emberlin, J. (2000). Pollen dispersal in the crops Maize (Zea mays), Oil seed rape (Brassica napus ssp oleifera), potatoes (Solanum tuberosum), sugar beet (Beta vulgaris ssp. Vulgaris) and wheat (Triticum aestivum), evidence for publications, a report for the soil association from the national pollen research unit, university college, Worcester, WR2 6AJ. Available at: http://www.soilasso ciation.org. Accessed Jan 2010. Tynan, J. L., Williams, M. K., & Conner, A. J. (1990). Low frequency of pollen dispersal from a eld trial of transgenic potatoes. Journal of Genetics and Breeding, 44, 303306. Wagner, D. B., & Allard, W. B. (1991). Pollen migration in predominantly self-fertilizing plants: Barley. The Journal of Heredity, 82, 392404. Yu, Q., Navajas-Perez, R., Tong, E., Robertson, J., Moore, P. H., & Paterson, A. H. (2008). Recent origin of dioecious and gynodioecious Y chromosomes in papaya. Tropical Plant Biology, 1, 4957.

123