臺灣博碩士論文加值系統

本論文的目的在於藉由收集生命表資料以比較帶有柑桔黃龍病菌（Candidatus Liberibacter asiaticus, Las (+)）與健康無菌（Las (-)）之亞洲柑桔木蝨（Diaphorina citri Kuwayama）生活史特徵與族群介量之差異。生活史觀察後死亡的蟲體分別以黃龍病菌特殊引子對配合PCR法來檢測是否帶菌，收集資料再以兩性生命表分析法進行分析。結果顯示帶菌的木蝨平均壽命比健康無菌木蝨有稍長，分別為35.2及33.4天。黃龍病菌會使某些木蝨若蟲發育不正常或羽化失敗，五齡若蟲期甚至會延長至一個月之久（最大值為40天）且無法發育到成蟲期，但是外部形態與正常發育的若蟲無明顯差異。而有些可成功發育到成蟲期的帶菌木蝨在羽化後可能會具有扭曲畸形的翅，這些情況都不曾發生在健康無菌的木蝨上。在每日產卵模式觀察中，木蝨族群傾向於在陽光較弱的早晨及下午產卵，夜間較少產卵。此外，帶有柑桔黃龍病菌的雌蟲會比健康無菌者具有較長之繁殖前期（分別為10.5 及5.7天），較晚的首次繁殖日期（自出生後算起，分別為27.6 及 21.9 天）及較低的產卵力(分別為18.6 及 29.6 粒卵/♀）。在此試驗中，無論帶菌與否，雌蟲都傾向於在產卵期之前段產下絕大部分的卵。而在兩者之族群介量方面，帶菌的木蝨雌蟲平均世代時間（T）與族群倍增時間（Dt）均長於健康無菌者（依序為29.09 vs. 22.87與10.35vs. 6.36天）。就內在增殖率（rm）而言，健康無菌者為0.1112 d-1為帶菌者（0.0689 d-1）之1.61倍，淨繁殖率（R0）為1.70倍（12.27 vs.7.22 offspring/female/generation），而終極增長率（λ）則為1.04倍(1.1176 vs. 1.0712 d-1)。在帶菌與健康無菌的木蝨族群穩定齡期分佈中，未成熟期均占極大比例，分別為84.25及88.58%。在帶菌木蝨族群中，具有傳播黃龍病菌能力的四、五齡若蟲期及成蟲期共占了48.03%，遠高於無菌族群之34.08%。綜合上述，黃龍病菌的存在確實會影響柑桔木蝨生活史特徵與族群介量。不同的病菌數量會造成木蝨不同的生理反應，而此數量又與木蝨首次獲菌量有關。當田間罹病病株率高時，木蝨帶菌率會隨之增加。故在黃龍病的綜合防治方面，清除田間病株與加強若蟲的防治均不容忽視。

This research was conducted to compare the differences in the life history traits and population parameters between Asian citrus psyllids (Diaphorina citri Kuwayama) with or without citrus Huanglongbing (HLB) pathogen (Candidatus Liberibacter asiaticus) (Las). The life table data were collected and then subjected to the age-stage, two-sex life table analysis. The polymerase chain reaction (PCR) was used to monitor the pathogen at the death of each psyllid. The results showed that the life span of Las-infected (Las (+)) psyllids were relatively longer than those of Las-free (Las (-)) ones, with 35.2 and 33.4 d. The Las appeared to cause psyllids to develop abnormally or emerge unsuccessfully. It also occasionally prolonged the period of 5th instar up to 40 d, resulting in their failure to reach adulthood. Some Las (+) adults even had abnormal wings after emergence. These abnormalities had never been observed in the Las (-) adults. Adult females had the preference of laying eggs during their early reproductive period, and daily oviposition pattern occurred primarily in two periods, one morning (7:30-9:30 a.m.) and the other late afternoon (3:30-5:30 p.m.). Comparing to Las (-) females, the Las (+) females also had longer pre-oviposition period (10.5 vs. 5.7 d), late first reproduction age counted since birth (27.6 vs. 21.9 d), and lower fecundity (18.6 vs. 29.6 eggs/♀). When comparing the population parameters between psyllids with or without being infected with the pathogen, the Las (+) psyllids had a longer mean generation time (T) (29.09 vs. 22.87 d) and doubling time (Dt) (10.35 vs. 6.36 d). The intrinsic rate of increase (rm) of the Las (-) female (0.1112 d-1) was 1.61 times higher than that of the Las (+) female (0.0689 d-1); the net reproductive rate (R0) was 1.70 times higher (12.27 vs. 7.22 offspring/female/generation); and the finite rate of increase (λ) was 1.04 times higher (1.1176 vs. 1.0712 d-1). In the stable stage distribution of Las (+) and Las (-) psyllids, the total percentage of immature was 84.25% and 88.58%, respectively. And the total percentage of the Las-transmitting individuals, including the 4th, 5th instar nymphs and adults, of Las (+) psyllids was 48.03%, which was higher than that of the Las (-) ones (34.08%). It is concluded that Las amounts found in psyllids would affect their life history traits and population parameters. Different amounts of Las found in a psyllid would cause different physiological responses. These Las amounts were closely related to the amount of Las acquired in the initial acquisition period. Furthermore, when the proportion of the infected citrus trees increased, the Las (+) psyllids would also increase. Therefore, removing and destroying infected citrus plants and improving the control of psyllid nymphs were essential to an effective field management of HLB.

Acknowledgements (Chinese) i
Abstract (Chinese) ii
Abstract (English) iv
Table of Contents vii
List of Tables ix
List of Figures x
List of Appendices xi
Introduction 1
Materials and Methods 5
1. Psyllid stock colonies 5
2. Host plant sources 5
3. Life table study 6
4. Daily reproductive schedule of females 7
5. Detection of the pathogen 7
6. Comparison of Las amount in the psyllid vectors 9
7. Life table analysis 9
8. Statistical analysis 10
Results 11
1. Seasonal viruliferous rate of psyllids 11
2. The age-stage, two-sex life table 11
2.1. Nymphal stage 11
2.1.1. Las (+) psyllid 11
2.1.2. Las (-) psyllid 12
2.2. Adult longevity and population parameters 13
2.2.1. Las (+) psyllid 13
2.2.2. Las (-) psyllid 14
2.2.3. Comparison between Las (+) and Las (-) psyllid 15
3. Influence of Las amounts in the psyllid vector 15
4. Reproductive schedule of females 16
Discussion and Conclusions 19
References 24
Tables 32
Figures 35
Appendices 49

APHIS news release. 2005. U. S. Department of Agriculture and Florida Department of Agriculture confirm detection of citrus greening. http:// www.aphis.usda.gov./lpa/news/2005/09/greening-pp8.doc.
Birch, L. C. 1948. The intrinsic rate of natural increase of an insect population. J. Anim. Ecol. 17: 15-26.
Bove, J. M., N. M. Chau, H. M. Trung, J. Bourdeaut, and M. Garnier. 1996. Huanglongbing (greening) in Viet Nam: Detection of Liberobacter asiaticum by DNA-hybridization with probe In 2.6 and PCR-amplification of 16S ribosomal DNA pp. 258-266. In: J. V. da Graca, P. Moreno, and R. K. Yokomi, eds. Proc. 13th Conference of the International Organization of Citrus Virologists (IOCV). University of California.
Capoor, S. P., D. G. Rao, and S. M. Viswanath. 1967. Diaphorina citri Kuwayama, a vector of the Greening disease of citrus in India. Indian J. Agr. Sci. 37: 572-576.
Chen, C. N. 1990. Integrated control of citrus pests in Taiwan. Fruits 45: 33-36.
Chen, M. H., T. Miyakawa, and C. Matsui. 1973. Citrus likubin pathogens in salivary glands of Diaphorina citri. Phytopathology 63: 194-195.
Chi, H. 1988. Life-table analysis incorporating both sexes and variable development rates among individuals. Environ. Entomol. 17: 26-34.
Chi, H. 2005. TWOSEX-MSChart: A computer program for the age-stage, two-sex life table analysis. http://140.120.197.173/Ecology/Download/Twosex-MSChart. zip.
Chi, H., and H. Liu. 1985. Two new methods for the study of insect population ecology. Bull. Inst. Zool., Academia Sinica 24: 225-240.
Chi, H., and T. C. Yang. 2003. Two-sex life table and predation rate of Propylaea japonica Thunberg (Coleoptera: Coccinellidae) fed on Myzus persicae (Sulzer) (Homoptera: Aphididae). Environ. Entomol. 32: 327-333.
Chiang-Lin, H. 2005. Multiplication and distribution of Candidatus Liberibacter asiaticus, the bacterium causing citrus huanglongbing, in the Asian citrus psyllid (Diaphorina citri Kuwayama). Master Thesis, National Taiwan University. (in Chinese). pp. 15-18.
Chien, C. C. 1992. Bionomics of Tamarixia radiate (Waterston): an ectoparasitoid of Diaphorina citri Kuwayama in Taiwan. Doctoral Dissertation, National Taiwan University. (in Chinese). pp. 44-48.
Da Graca, J. V. 1991. Citrus greening disease. Annu. Rev. Phytopathol. 29: 109-136.
Etienne, J., S. Quilici, D. Marival, and A. Franck. 2001. Biological control of Diaphorina citri (Hemiptera: Psyllidae) in Guadeloupe by imported Tamarixia radiate (Hymenoptera: Eulophidae). Fruits 56: 307-315.
Fung, Y. C., and C. N. Chen. 2006. Effects of temperature and host plant on population parameters of the citrus psyllid (Diaphorina citri Kuwayama). Formosan Entomol. 26: 109-123. (in Chinese)
Garnier, M., and J. M. Bove. 1983. Transmission of the organism associated with citrus greening disease from sweet orange to periwinkle by dodder. Phytopathology 73: 1358-1363.
Grafton-Cardwell, E. E., K. E. Godfrey, M. E. Rogers, C. C. Childers, and P. A. Stansly. 2006. Asian citrus psyllid. University of California Exotic and Invasion Pests and Disease Research Program pp. 1-8.
Halbert, S. E., and K. L. Manjunath. 2004. Asian citrus psyllids (Sternorrhyncha: Psyllidae) and greening disease of citrus: A literature review and assessment of risk in Florida. Florida Entomol. 87: 330-353.
Halbert, S. E., and C. A. Nunez. 2004. Distribution of the Asian citrus psyllid, Diaphorina citri Kuwayama (Rhynchota: Psyllidae) in the Caribbean Basin. Florida Entomol. 87: 401-402.
Halbert, S. E., X. A. Sun, and W. N. Dixon. 2000. Asian citrus psyllid and citrus greening disease. Citrus Ind. 81(5): 22-24.
Hoy, M. A., A. Jeyaprakash, and R. Nguyen. 2001. Long PCR is a sensitive method for detecting Liberobacter asiaticum in parasitoids undergoing risk assessment in quarantine. Biol. Control 22: 278-287.
Hoy, M. A., R. Nguyen, and A. Jeyaprakash. 1999. Classical biological control of Asian citrus psyllid. Citrus Ind. 80: 20-22.
Huang, A. L. 1987. Electronmicroscopical studies on the morphology and population dynamic of fastidious bacteria causing citrus likubin. Ph.D. Dissertation. National Taiwan University, Taiwan.
Huang, C. H. 1979. Distribution of likubin pathogen in likubin-affected citrus plants. Agric. Res. China (Taiwan) 28(1): 29-33. (in Chinese)
Huang, C. H., M. Y. Tsai, and C. L. Wang. 1984. Transmission of citrus likubin by a psyllid, Diaphorina citri. Agric. Res. China (Taiwan) 33(1): 65-72.
Hung, S. C. 2006. Ecology and vectorship of the citrus psyllid in relation to the prevalence of citrus huanglongbing. Doctoral Dissertation, National Taiwan University. (in Chinese). pp. 76-77, 81-86.
Hung, T. H., M. L. Wu, and H. J. Su. 1999. Development of a rapid method for the diagnosis of citrus greening disease using the polymerase chain reaction. J. Phytopathol. 147: 599-604.
Hung, T. H., M. L. Wu, and H. J. Su. 2001. Identification of the Chinese box orange (Severinia buxifolia) as an alternative host of the bacterium causing citrus Huanglongbing. Euro. J. Plant Pathol. 107: 183-189.
Hung, T. H., S. C. Hung, C. N. Chen, M. H. Hsu, and H. J. Su. 2004. Detection by PCR of Candidatus Liberibacter asiaticus, the bacterium causing citrus huanglongbing in vector psyllids: application to the study of vector-pathogen relationships. Plant Pathol. 53: 96-102.
Jagoueix, S., J. M. Bove, and M. Garnier. 1997. Comparison of the 16S/23S ribosomal intergenic regions of ‘Candidatus Liberobacter asiatium’ and ‘Candidatus Liberobacter africanum’, the two species associated with citrus huanglongbing (greening) disease. International System. Bacteriol. 47: 224-227.
Knapp, J. L., S. Halbert, R. Lee, M. Hoy, R. Clark, and M. Kesinger. 1998. The Asian citrus psyllid and citrus greening disease. Citrus Ind. 79(10): 28-29.
Lakra, R. K., Z. Singh, and W. S. Kharub. 1983. Population dynamics of citrus psylla, Diaphorina citri Kuwayama in Haryana. Indian J. Entomol. 45: 301-310.
Lee, R. F. 1996. Citrus greening disease and its vectors. Naranja 17: 258-260.
Lewontin, R. C. 1965. Selection for colonizing ability. pp. 77-91. In: H. G. Baker, and G. L. Stebbins, eds. The Genetics of Colonizing Species. Academic Press, New York.
Liu, Y. H., and J. H. Tsai. 2000. Effects of temperature on biology and life table parameters of the Asian citrus psyllid, Diaphorina citri Kuwayama (Homoptera: Psyllidae). Ann. Appl. Biol. 137: 201-206.
Martinez, A. L., D. M. Nora, and A. L. Armedilla. 1970. Suppression of symptoms of citrus greening disease in the Philippines by treatment with tetracycline antibiotics. Plant Disease Reporter 54: 1007-1009.
Matsumoto, T., M. C. Wang, and H. J. Su. 1961. Studies on likubin. pp. 121-125. In: Price, W. C., eds. Proc. 2nd Conference of the International Organization of Citrus Virologists (IOCV). University of Florida, Gainesville.
Mckenzie, C. L., and G. J. Puterka. 2004. Effect of sucrose octanoate on survival of nymphal and adult Diaphorina citri (Homoptera: Psyllidae). J. Econ. Entomol. 97: 970-975.
Michaud, J. P. 2000. The Asian citrus psyllid and its natural enemies. Citrus Ind. 81(8): 42-44.
Michaud, J. P. 2004. Natural mortality of Asian citrus psyllid (Homoptera: Psyllidae) in central Florida. Biol. Control 29: 260-269.
Miyakawa, T., and Z. X. Yuan. 1990. Citrus host range of greening disease. pp. 118-121. In: B. Aubert, S. Tontyaporn, and D. Buangsuwon, eds. Proc. 4th International Asia Pacific Conference on Citrus Rehabilitation. FAO-UNDP.
Pluke, R. W. H., A. Escribano, J. P. Michaud, and P. A. Stansly. 2005. Potential impact of lady beetles on Diaphorina citri (Homoptera: Psyllidae) in Puerto Rico. Florida Entomol. 88: 123-128.
Rubinstein, G., and H. Czosnek. 1997. Long-term association of tomato yellow leaf curl virus with its whitefly vector Bemisia tabaci: effect on the insect transmission capacity, longevity and fecundity. Gen. Virol. 78: 2683-2689.
Skelley, L. H., and M. A. Hoy. 2004. A synchronous rearing method for the Asian citrus psyllid and its parasitoids in quarantine. Biol. Control 29: 14-23.
Su, H. J., J. U. Cheon, and M. J. Tsai. 1986. Citrus greening (likubin) and some viruses and their control trials. pp. 142-147. In: Plant Virus Disease of Horticultural Crops in the Tropics and Subtropics. FFTC Book Series No.33, FFTC for the ASPAC Region, Taipei, Taiwan.
Tsai, J. H., and Y. H. Liu. 2000. Biology of Diaphorina citri (Homoptera: Psyllidae) on four host plants. J. Econ. Entomol. 93: 1721-1725.
Tsai, J. H., J. J. Wang, and Y. H. Liu. 2000. Sampling of Diaphorina citri (Homoptera: Psyllidae) on orange Jessamine in southern Florida. Florida Entomol. 83: 446-459.
Tsai, J. H., J. J. Wang, and Y. H. Liu. 2002. Seasonal abundance of the Asian citrus psyllid, Diaphorina citri (Homoptera: Psyllidae) in southern Florida. Florida Entomol. 85: 446-451.
Van Den Berg, M. A. 1986. Effects of citrus cultivars and reduced irrigation on availability of new growth for citrus psylla breeding. Fruits 41: 597-604.
Van Vuuren, S. P. 1993. Variable transmission of African greening to sweet orange. pp. 264-268. In: P. Moreno, J. V. da Graca, and L. W. Timmer, eds. Proc. 12th Conference of the International Organization of Citrus Virologists. University of California, Riverside.
Xu, C. F., Y. H. Xia, K. B. Li, and C. Ke. 1988. Further study of the transmission of citrus huanglongbing by a psyllid, Diaphorina citri Kuwayama. pp. 243-248. In: L. W. Timmer, S. M. Garnsey, and L. Navarro, eds. Proc. 10th Conference of the International Organization of Citrus Virologists. Riverside, CA.
Zou, M., and C. Y. Zhou. 2005. Advances in pathogen research and detection technique of huanglongbing. Plant Protection 31(3): 10-14. (in Chinese)