蜜蜂球囊菌遗传转化体系构建及其致病性的分子机理研究
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摘要
蜜蜂是世界上最重要的经济昆虫,在世界上只要有花和花粉的地方就有蜜蜂。然而在生态上如此重要的昆虫却遭受着各种各样疾病的侵染,这些疾病可能由多种病原物引起,如真菌就是其中一种重要的病原微生物。对真菌病害的致病性机理的研究一直以来都是研究的热点。蜜蜂白垩病是由蜜蜂球囊菌(Ascosphaera apis)引起的,主要侵染蜜蜂的幼虫。研究表明,白垩病可致使蜜蜂蜂蜜产量减产5%-37%,使蜜蜂幼虫产生80%的死亡率。近些年来,由于蜜蜂白垩病发病较重,人们对杀菌剂使用较乱,导致蜂产品污染严重。再加上蜜蜂的转地饲养,蜜蜂长距离的运输又导致了蜜蜂白垩病的传播具有全球性。更为严重的是,引起蜜蜂白垩病的致病菌球囊菌的分生孢子可以在蜂蜜、花粉、蜂机具及发病蜂场的土壤中存活15年以上,当条件适宜其生长时,分生孢子会从环境中萌发出来,再次传播,使得该病很难根除。尽管科研人员提出了许多防控蜜蜂白垩病的技术措施,然而至目前为止,对蜜蜂白垩病的防治还是非常困难的。鉴于此,本研究采用分子生物学方法,利用限制性内切酶介导的基因整合技术构建了蜜蜂球囊菌遗传转化体系,获得了球囊菌致病性降低突变体,对蜜蜂球囊菌致病的分子机理进行了初步的探讨。
1.蜜蜂球囊菌原生质体制备及其再生
构建蜜蜂球囊菌REMI转化体系首先要制备出产量高再生率高的原生质体。本实验对影响蜜蜂球囊菌菌株原生质体制备及再生的因子进行了系统的研究,同时对蜜蜂球囊菌原生质体的释放及再生过程进行了显微观察。结果表明,采用液体培养基进行24hrs培养的蜜蜂球囊菌菌体,在28℃条件下,应用50mg/mL的崩溃酶,经4hrs酶解所制备的原生质体释放量最大,达到98.36×105/mL。在上述条件下,采用0.8mol/L柠檬酸与NaCl的混合液作为稳渗剂,原生质体的再生率也较高,达到51.06%。蜜蜂球囊菌以菌丝断裂方式释放原生质体,原生质体再生时表现为原生质体先是突出,其后延长,并最终发育成为正常菌丝。
2. REMI转化子的获得及其验证
采用REMI方法获得了11个转化子。经过PCR. ClustalW多序列比对及Southern blot方法验证,所获得突变体含有目的基因的插入,由此证明上述转化子为蜜蜂球囊菌突变菌株。
3.蜜蜂球囊菌REMI突变子致病性鉴定
通过幼虫接种实验,对所获得的REMI突变菌株进行了致病力进行了检测。结果表明,所获得的突变体菌株同原始菌株比较,致病力差异显著(p<0.01),接种后的第5天野生型菌株引起幼虫发病率达19.9%,病情指数达33.5,M4和M8的致病性明显小于野生型菌株,幼虫发病率仅为16.8%和14.3%,病情指数为28.9和23.7。同时,突变体菌株在侵染蜜蜂幼虫后,蜜蜂发病过程及幼虫死亡率均较野生型有较大差异。
Honeybees are one of the most well-known economically beneficial social insects which can be kept almost anywhere in the globe as long as there are flowering plants that produce nectar and pollen. These insects are accomplishing about80%of all crop insect pollinations. However, this ecologically important social insect is suffering from various diseases caused by various pathogens, like fungal diseases. Of course, fungi are economically important organisms whose pathogenicity continues to be the focus of extensive research. Among the fungal diseases, chalkbrood is the one known to be an invasive mycosis in honeybees, caused by Ascosphaera apis, that exclusively affects honeybee brood. The disease has been reported to cause about5%-37%reduction in honey production and80%brood death. Consequently, the severity of the disease, these days, has increased the use of pesticides and frequent long-distance transportation of colonies which is believed to provide an additional opportunity for A. apis infection globally. Furthermore, the fact that A. apis is so widespread in the globe and its viable spores can be found in stored honey, pollen, pollen capsules/tablets, used hive components, used beekeeping tools and equipments, and possibly in soil around infected apiaries for more than15years, makes the possibility of its eradication most unlikely. Here, even though a broad range of experimental works have been conducted safe control of the disease has been noticed to be difficult. Thus, genetic engineering works for better manipulation of the fungus, molecular investigation of pathogenesis and possible identification of strategies to control the disease shall take current issue of research. With this, this work was conducted with the objectives of understanding and developing an efficient and reproducible technique for the ultimate purpose of making this fungal species amenable to genetic studies and transformation. Furthermore, this work was aimed to study the pathogenicity of restricted enzyme mediated integration engineered A. apis mutants in honeybee(Apis melifera L.) larvae. Consequently, as restricted enzyme-mediated integration enables the investigation of the infectivity of transformed pathogens to host organisms, this work has utilized protoplast isolation and transformation techniques as a tool to understand this fungus further to meet set objectives. According to the results obtained, the fungus showed varying responses in terms of yield, size, and regeneration rates on different mediums, and osmotic stabilizer combinations. In this way, it was confirmed that younger incubation time has yielded better mycelium suitable for highest protoplast isolation while use of driselase was the best enzyme treatment, yielded about98.36×105mL-1of protoplasts. Young aged and exponentially growing mycelial culture provided the highest protoplast formation with higher protoplast viability. Use of citric acid monohydrate with NaCl as an osmotic stabilizer supported a51.06%viable protoplasts to regenerate and able to form mycelia colonies, while a combination of4hrs enzymolysis time with this osmotic stabilizer have supported53.06%protoplast regeneration. Following the successful protoplast isolation and restricted enzyme-mediated integration, PCR, ClustalW multiple sequence alignment, and Southern blot analyses confirmed the successful integration of a foreign DNA, as an insert, to the host chromosome targeting to cause mutation. An in-vitro bioassay experiment included in this work has confirmed that all engineered mutants had notable differences in pathogenicity among themselves and with the wild strain to the honeybee larvae (p<0.01). Disease infectivity data confirmed that the original (wild type) fungus caused the maximum (19.9%) amount of larval death starting from the5th day post inoculation, and its disease index was recorded to be33.5. Even though the mortality of engineered mutants was not significantly augmented compared to that of the wild strain, mutants4and8had16.8%and14.3%larval death with28.9and23.7of disease index, respectively. More specifically, transformed A. apis mutants were relatively less pathogenic to in vitro-reared honeybee larvae than those of the wild-type. In addition, higher larval death and faster mummification was detected in the wild-type too. Thus, the pathogenicity differences among strains, even in the natural conditions, have magnified the significance of further investigations on the reasons for variations among transformants and their wild types. Even if we have studied this fungus for a while, control of this fungus is still a mystery. Thus, we, generally, recommend that further understanding on this fungus and different investigations shall be carried out as a part of future important activities.
1.蜜蜂球囊菌原生质体制备及其再生
构建蜜蜂球囊菌REMI转化体系首先要制备出产量高再生率高的原生质体。本实验对影响蜜蜂球囊菌菌株原生质体制备及再生的因子进行了系统的研究,同时对蜜蜂球囊菌原生质体的释放及再生过程进行了显微观察。结果表明,采用液体培养基进行24hrs培养的蜜蜂球囊菌菌体,在28℃条件下,应用50mg/mL的崩溃酶,经4hrs酶解所制备的原生质体释放量最大,达到98.36×105/mL。在上述条件下,采用0.8mol/L柠檬酸与NaCl的混合液作为稳渗剂,原生质体的再生率也较高,达到51.06%。蜜蜂球囊菌以菌丝断裂方式释放原生质体,原生质体再生时表现为原生质体先是突出,其后延长,并最终发育成为正常菌丝。
2. REMI转化子的获得及其验证
采用REMI方法获得了11个转化子。经过PCR. ClustalW多序列比对及Southern blot方法验证,所获得突变体含有目的基因的插入,由此证明上述转化子为蜜蜂球囊菌突变菌株。
3.蜜蜂球囊菌REMI突变子致病性鉴定
通过幼虫接种实验,对所获得的REMI突变菌株进行了致病力进行了检测。结果表明,所获得的突变体菌株同原始菌株比较,致病力差异显著(p<0.01),接种后的第5天野生型菌株引起幼虫发病率达19.9%,病情指数达33.5,M4和M8的致病性明显小于野生型菌株,幼虫发病率仅为16.8%和14.3%,病情指数为28.9和23.7。同时,突变体菌株在侵染蜜蜂幼虫后,蜜蜂发病过程及幼虫死亡率均较野生型有较大差异。
Honeybees are one of the most well-known economically beneficial social insects which can be kept almost anywhere in the globe as long as there are flowering plants that produce nectar and pollen. These insects are accomplishing about80%of all crop insect pollinations. However, this ecologically important social insect is suffering from various diseases caused by various pathogens, like fungal diseases. Of course, fungi are economically important organisms whose pathogenicity continues to be the focus of extensive research. Among the fungal diseases, chalkbrood is the one known to be an invasive mycosis in honeybees, caused by Ascosphaera apis, that exclusively affects honeybee brood. The disease has been reported to cause about5%-37%reduction in honey production and80%brood death. Consequently, the severity of the disease, these days, has increased the use of pesticides and frequent long-distance transportation of colonies which is believed to provide an additional opportunity for A. apis infection globally. Furthermore, the fact that A. apis is so widespread in the globe and its viable spores can be found in stored honey, pollen, pollen capsules/tablets, used hive components, used beekeeping tools and equipments, and possibly in soil around infected apiaries for more than15years, makes the possibility of its eradication most unlikely. Here, even though a broad range of experimental works have been conducted safe control of the disease has been noticed to be difficult. Thus, genetic engineering works for better manipulation of the fungus, molecular investigation of pathogenesis and possible identification of strategies to control the disease shall take current issue of research. With this, this work was conducted with the objectives of understanding and developing an efficient and reproducible technique for the ultimate purpose of making this fungal species amenable to genetic studies and transformation. Furthermore, this work was aimed to study the pathogenicity of restricted enzyme mediated integration engineered A. apis mutants in honeybee(Apis melifera L.) larvae. Consequently, as restricted enzyme-mediated integration enables the investigation of the infectivity of transformed pathogens to host organisms, this work has utilized protoplast isolation and transformation techniques as a tool to understand this fungus further to meet set objectives. According to the results obtained, the fungus showed varying responses in terms of yield, size, and regeneration rates on different mediums, and osmotic stabilizer combinations. In this way, it was confirmed that younger incubation time has yielded better mycelium suitable for highest protoplast isolation while use of driselase was the best enzyme treatment, yielded about98.36×105mL-1of protoplasts. Young aged and exponentially growing mycelial culture provided the highest protoplast formation with higher protoplast viability. Use of citric acid monohydrate with NaCl as an osmotic stabilizer supported a51.06%viable protoplasts to regenerate and able to form mycelia colonies, while a combination of4hrs enzymolysis time with this osmotic stabilizer have supported53.06%protoplast regeneration. Following the successful protoplast isolation and restricted enzyme-mediated integration, PCR, ClustalW multiple sequence alignment, and Southern blot analyses confirmed the successful integration of a foreign DNA, as an insert, to the host chromosome targeting to cause mutation. An in-vitro bioassay experiment included in this work has confirmed that all engineered mutants had notable differences in pathogenicity among themselves and with the wild strain to the honeybee larvae (p<0.01). Disease infectivity data confirmed that the original (wild type) fungus caused the maximum (19.9%) amount of larval death starting from the5th day post inoculation, and its disease index was recorded to be33.5. Even though the mortality of engineered mutants was not significantly augmented compared to that of the wild strain, mutants4and8had16.8%and14.3%larval death with28.9and23.7of disease index, respectively. More specifically, transformed A. apis mutants were relatively less pathogenic to in vitro-reared honeybee larvae than those of the wild-type. In addition, higher larval death and faster mummification was detected in the wild-type too. Thus, the pathogenicity differences among strains, even in the natural conditions, have magnified the significance of further investigations on the reasons for variations among transformants and their wild types. Even if we have studied this fungus for a while, control of this fungus is still a mystery. Thus, we, generally, recommend that further understanding on this fungus and different investigations shall be carried out as a part of future important activities.
引文
Abebe J.W, Shufa X., Yingying H., Wei L., Zhanbao G., Fei X., Ting Z.,2013. Current developments in Ascosphaera apis (Maasen ex Claussen) L.S. Olive Spiltoir:the causative agent of Chalkbrood disease in Honeybees (Hymenoptera:Apidae). Journal of Food, Agriculture & Environment-JFAE 11 (3&4):2190-2200.
Aizen M., Garibaldi L., Cunningham S., and Klein A.,2008. Long-term global trends in crop yield and production reveal no current pollination shortage but increasing pollinator dependency. Current Biology 18:1572-1575.
Aizen M., and Harder L.D.,2009. The global stock of domesticated honey bees is growing slower than agricultural demand for pollination. Current Biology 19:915-918.
Aizen M.A., Lucas A.G., Saul A.C., and Alexandra M.K.,2009. How much does agriculture depend on pollinators? Lessons from long-term trends in crop production. Annals of Botany 103: 1579-1588.
Alonso J.M, Rey J., Puerta F., Hermoso de Mendoza J., Hermoso de Mendoza M., and Flores J.M., 1993. Enzymatic equipment of Ascosphaera apis and the development of infection by this fungus in Apis mellifera. Apidologie 24:383-390.
Amaya E., and Kroll K.,2010. Production of Transgenic Xenopus laevis by Restriction Enzyme Mediated Integration and Nuclear Transplantation. Journal of Visualized Experiments 42:doi: 10.3791/2010.
Anderson D.L., Gibbs A.J., and Gibson N.L.,1998. Identification and phylogeny of spore-cyst fungi(Ascosphaera spp.) using ribosomal DNA sequences. Mycological Research 102 (5): 541-547.
Anderson D.L, and Gibson N.L.,1998. New species and isolate of spore-cysts fungi (Plectomycetes: Ascosphaerales) from Australia, Austral, Systematic Botany 11:53-72
Anderson D., Giacon H., and Gibson N.,1997. Detection and thermal destruction of the chalkbrood fungus(Ascosphaera apis) in honey. Journal of Apiculture Research 36 (3-4):163-168.
Aronstein K.A, Murray K.D, and Saldivar E.,2010. Transcriptional responses in Honey Bee larvae infected with chalkbrood fungus. BMC Genomics 11:pp391.
Aronstein K.A, and Murray K.D.,2010. Chalkbrood disease in honey bees, Journal of Invertebrate Pathology 103:S20-S29.
Aupinel P., Fortini D., Dufour H., Tasei J.N., Michaud B., Odoux J.F., and Pham-Delegue M.H., 2005. Improvement of artificial feeding in a standard in vitro method for rearing Apis mellifera larvae. Bulletin of Insectology 58:107-111.
Aupinel P., Fortini D., Michaud B., Marolleau F., Tasel J.N., and Odoux J.F.,2007a. Toxicity of dimethoate and fenoxycarb to honey bee brood(Apis mellifera), using a new in vitro standardized feeding method. Pest Management Science 63:1090-1094.
Aupinel P., Fortini D., Michaud B., Medrzycki P., Padovani E., Przygoda D., Maus C., Charriere J.D., Kilchenmann V., Riessberger-Galle U., Vollmann J.J., Jeker L., Janke M., Odoux J.F., and Tasei J.N.,2009. Honey bee brood ring-test:method for testing pesticide toxicity on honey bee brood in laboratory conditions. Julius-Kuhn-Archiv 423:96-102.
Aupinel P., Medrzycki P., Fortini D., Michaud B., Tasei J.N., and Odoux J.F.,2007b. A new larval in vitro rearing method to test effects of pesticides on honey bee brood. Redia 90:91-94.
Bailey L.,1963. Infectious Diseases of the Honeybee, Land Books Ltd. London, pp176.
Bailey L.,1967. The effect of temperature on the pathogenicity of the fungus Ascosphaera apis, for brood of the honey bee, Apis mellifera. In Proceedings of the International Cooloquium on Insect Pathology and Microbial Control (P.A. vab der Laan, Ed. pp162-167.
Bailey L., and Ball B.V.,1991. Honey Bee Pathology. Academic Press, London, pp53-63; pp154-158
Bailey L., and Fernando, E.F.W.,1972. Effects of sacbrood virus on adult honey bees. Annals of Applied Biology 72:27-35.
Bakhiet N., and Stahly D.P.,1985. Ultrastructure of Sporulating Bacillus larvae in a Broth Medium. Applied and Environmental Microbiology 50(3):690-692.
Balhareth H.M., Alqarni, A.S. and Owayss, A.A.,2012. Comparison of hygienic and grooming behaviors of indigenous and exotic honeybee, Apis mellifera races in central Saudi Arabia. International Journal of Agriculture and Biology 14:1005-1008.
Bamford S.,1987. Studies on the infection of honeybee larvae with Ascosphaera apis. PhD Thesis, Plymouth Polytechnic, United Kingdom.
Bamford S., and Heath L.A.F.,1989. The effects of temperature and pH on the germination of spores of the Chalkbrood fungus,A scosphaera apis. Journal of Apicultural Research,28: 36-40.
Barnett E.A., Charlton A.J., and Fletcher M.R.,2007. Incidents of bee poisoning with pesticides in the United Kingdom 1994-2003, Pest Management Sciences 63:1051-1057
Benedek P.,1985. Economic importance of honeybee pollination of crops at the national level in Hungary, Proceedings of the 29th international beekeeping congress, Budapest, Hungary, Apimondia, pp286-289.
Bills S.N., Richter, D.L., and Podila, GK.,1995. Genetic transformation of the ectomycorrhizal fungus Paxillus involutus by particle bombardment. Mycological Research 99:557-561
Bissett J., Duke G, and Goettel M.,1996. Ascosphaera acerosa species nov. Isolated from the alfaalfa leafcutting bee, with a key to the species of Ascosphaera, Mycologia 88 (5):797-803
Bissett J.,1988. Contribution towards a monograph of the genus Ascosphaera. Canadian Journal of Botany 66:2541-2560.
Blochlinger K., and Diggelmann H.,1984. Hygromycin B Phosphotransferase as a Selectable Marker for DNA Transfer Experiments with Higher Eucaryotic Cells. Molecular Cell Biology 4(12):2929-2931.
Borum A.E., and Ulgen M.,2008. Chalkbrood(Ascosphaera apis) infection and fungal agents of honey bees in North-West Turkey. Journal of Apicultural Research 47(2):170-171
Bruun-Jensen A., Bo V.P., and Jorgen E.,2009. Differential susceptibility across honey bee colonies in larval chalkbrood resistance, Apidologie 40:524-534.
Camazine S.,1989. Beekeeping in South Korea. Bee World 70:66-74.
Campano F., Flores J.M., Puerta F., Ruiz J.A., and Ruz J.M.,1999. Fungal diseases of the honeybee (Apis mellifera L.). In:Colin M.E. (ed.), Ball B.V. (ed.), Kilani M. (ed.). Bee disease diagnosis. Zaragoza:CIHEAM, (Options Mediterraneennes:Serie B. Etudes et Recherches; n.25), pp61-68.
Carrera P., Sommaruga A., and Vailati G.,1987. The development of Ascosphaera apis within larvae of Apis mellifera ligustica. Journal of Apiculture Research 26(1):59-63.
Case M.E., Schweizer M., Kushner S.R., Giles N.H.,1979. Efficient transformation of Neurospora crassa by utilizing hybrid plasmid DNA. Proc. Natural Academic Seinces, USA.76: 5259-5263.
Chen R., and Jeffrey J.M.,1993. Production of Fungal Protoplasts from selected wood-degrading fungi. Oregon State University, Wood and Fiber Science 25 (1):61-65.
Chen Y.P. and Siede R.,2007. Honey bee viruses. Advances in Virus Research 70:33-80.
Chorbinski P., and Rypula K.,2003. Studies on the morphology of strains Ascosphaera apis isolated from chalkbrood disease of the honey bees. Veterinary Medicine 6 (2):1-12.
Chorbinski P.,2004. Identification of Ascosphaera apis strains using ribosomal DNA sequences. Medycyna Weterynaryjna 60(2):190-192.
Chung K.R., Ehrenshaft M., Wetzel D.K., and Daub M.E.,2003. Cercosporin-deficient mutants by plasmid tagging in the asexual fungus Cercospora nicotianae. Molecular Genetics and Genomics270:103-113.
Cocking E.C.,1960. A method for the isolation of plant protoplasts and vacuoles; Nature (London) 187:927-929.
Cocking E.C.,1979. All sorts of protoplasts; Nature,281:180-181. Davis BD,1985. Factors affecting protoplast isolation. In:Fungal protoplasts:Application in biochemistry and genetics (Peberdy JF, Ferenczy L eds.), Marcel Dekker, New York, pp45-72.
Cordoni G.,2011. Epidemiology and taxonomy of honey bee viruses in England and Wales. PhD thesis. University of Surrey.
Cordoni G., and Spagnuolo M.L.,2007. Development of an experimental GIS for bee-keeping in the Marche Region, Italy. Veterinaria Italiana 43(3):431-436.
Cornman R.S., Bennett A.K., Murray K.D., Evans J.D., Elsik C.G., and Aronstein K.,2012. Transcriptome analysis of the honey bee fungal pathogen, Ascosphaera apis:implications for host pathogenesis. BMC Genomics 13:pp285.
Cove D.J.,1979. The uses of isolated protoplasts in plant genetics. Heredity 43 (3):295-314.
Crailsheim K., Brodschneider R., Aupinel P., Behrens D., Genersch E., Vollmann J., and Riessberger-Galle U.,2013. Standard methods for artificial rearing of Apis mellifera larvae. Journal of Apiculture Research 52(1):1-15.
Crane E.,1991. Apis species of tropical Asia as pollinators and some rearing methods for them. Acta Horticulture 288:29-48.
Crane E.,1999. The World History of Beekeeping and Honey Hunting. New York:Routledge.
Cremer S., Jiirgen H., Alexandra S., Daniel J.C.K., and Line V.U.,2010. Rapid anti-pathogen response in ant societies relies on high genetic diversity. Proceedings of the Royal Society B: Biological Sciences 277:2821-2828
Cummings W.J., Celerin M., Crodian J., Brunick L.K., and Zolan M.E.,1999. Insertional mutageneses in Coprinus cinereus:use of a dominant selectable marker to generate tagged, sporulation-defective mutants. Current Genetics 36:371-382.
Cundliffe E.,1968. Preparation and Some Properties of Active Protoplasts of Bacillus megaterium, Journal of General Microbiology 53:425-430.
Davey M.R., Anthony P., Power J.B., and Lowe K.C.,2005. Plant protoplasts:status and biotechnological Perspectives. Biotechnological Advances 23:131-171.
Davis C., and Ward W.,2003. Control of chalkbrood disease with natural products:a report for the RIRDC. Publication No.03/107, Kingston, ACT, AU, ppl-23.
Deodikar G.B., Ghatge A.L., Phadke R.P., Mahindre D.B., Kshirsagar K.K., Muvel S., and Thakar S.S.,1977. Nesting behaviour of Indian honeybees. Ⅲ. Nesting behaviour of Apis dorsata. Indian Bee Journal 39:1-12.
Desalegn B.,2006. The Occurrence of Chalk Brood (Ascosphaera apis):A Honeybee (A. mellifera L.) Disease in West Shoa, Ethiopia, Ethiopian Journal of Animal Production 6(1):1-8.
Desneux N., Decourtye A., and Delpuech J.M.,2007. The sub lethal effects of pesticides on beneficial arthropods. Annual Review of Entomology 52:81-106.
Eguchi F., Renato G.R., Tomoaki I., and Miyato H.,1998. Regeneration of protoplasts from hyphal strands of Volvariella volvacea Journal of Wood Science 44(5):401-407.
Eguchi F., Tashiro M., Suzuki T., and Higaki M.,1990. Preparation and regeneration of protoplasts of edible mushrooms. Mokuzai Gakkaishi 36:232-240.
Engel M.S.,1999. "The taxonomy of recent and fossil honey bees (Hymenoptera:Apidae:Apis)". Journal of Hymenoptera Research 8:165-196.
Engel M.S., Hinojosa-Diaz I.A., and Rasnitsyn A.P.,2009. "A honey bee from the Miocene of Nevada and the biogeography of Apis (Hymenoptera:Apidae:Apini)". Proceedings of the California Academy of Sciences 60 (3):23-38.
European Synchrotron Radiation Facility,2012. First ever record of insect pollination from 100 million years ago. ScienceDaily. Retrieved October 8,2013, from http://www.sciencedaily.com/releases/2012/05/120514153113.htm.
Evans J.D., Aronstein K., Chen Y.P., Hetru C., Imler J.L., Jiang H., Kanost M., Thompson G.J., Zou Z., and Hultmark D.,2006. Immune pathways and defence mechanisms in honey bees Apis mellifera. Insect Molecular Biology 15(5):645-656.
Evans J.D., and Spivak M.,2010. Socialized medicine:Individual and communal disease barriers in honey bees. Journal of Invertebrate Pathology 103:S62-S72.
Feng F.,2000. Infectious diseases in honeybee and the control methods. Journal of bee (7):pp19
Fincham J.R.S.,1989. Transformation in fungi. Microbiological Review 53:148-170.
Flores J.M., Gutierrez I., and Espejo R.,2005b. The role of pollen in chalkbrood disease in Apis mellifera:transmission and predisposing conditions. Mycologia 97:1171-1176.
Flores J.M., Gutierrez I., and Puerta F.,2004a. A comparison of methods to experimentally induce chalkbrood disease in honey bees. Spanish Journal of Agricultural Research 2 (1):79-83.
Flores J.M., Gutierrez I., and Puerta F.,2004b. Oxytetracycline as a predisposing condition for chalkbrood in honeybee. Veterinary Microbiology 103:195-199.
Flores J., Spivak M., and Gutierrez I.,2005a. Spores of Ascosphaera apis contained in wax foundation can infect honeybee brood. Veterinary Microbiology 108:141-144.
Flores J.M., Ruiz J.A., Ruz J.M., Puerta F., Bustos M., Padilla F., and Campano F.,1996. Effect of temperature and humidity of sealed brood on chalkbrood development under controlled conditions. Apidologie 27:185-192.
Flowers J.L., and Vaillancourt L.J.,2005. Parameters affecting the efficiency of Agrobacterium tumefaciensmediated transformation of Colletotrichum graminicola. Current Genetics 48: 380-388.
Fries I., and Camazine S.,2001. Implications of horizontal and vertical pathogen transmission for honey bee epidemiology. Apidologie 32(3):199-214.
Fries I., and Lindstrom A.,2010. Breeding disease resistant honeybees, Swedish University of Agricultural Sciences, Uppsala, Sweden. pp21.
Fujimoto D., Shi Y., Christian D., Mantaguihan J.B., and Leung H.,2002 Tagging quantitative loci controlling pathogenicity in Magnaporthe grisea by insertional mutagenesis. Physiological and Molecular Plant Pathology 61:77-88.
Fungaro M.H.P., Rech E., Muhlen GS., Vainstein M.H., Pascon R.C., de Queiroz M.V., Pizzirani-Kleiner A.A., and de Azevedo J.L.,1995. Transformation of Aspergillus nidulans by microprojectile bombardment on intact conidia. FEMS Microbiological Letters 125:293-298.
Gallai N., Salles J.M., Settele J., and Vaissie're B.E.,2009. Economic valuation of the vulnerability of world agriculture confronted with pollinator decline. Ecological Economics 68:810-821.
Gao C., Xue Y., and Ma Y,2011. Protoplast Transformation of Recalcitrant Alkaliphilic Bacillus sp. with Methylated Plasmid DNA and a Developed Hard Agar Regeneration Medium; PLoS ONE 6(11):e28148. doi:10.1371/journal.pone.0028148
Genersch E., Ashiralieva A., and Fries I.,2005. Strain- and genotype-specific differences in virulence of Paenibacillus larvae subsp. larvae, a bacterial pathogen causing American foulbrood disease in honey bees. Applied Environmental Microbiology, pp7551-7555. http://dx.doi.org/10.1128/AEM.71.11.7551-7555.2005.
Genersch E., Forsgren E., Pentikainen J., Ashiralieva A., Rauch S., Kilwinski J., and Fries I.,2006. Reclassification of Paenibacillus larvae subsp. pulvifaciens and Paenibacillus larvae subsp. larvae as Paenibacillus larvae without subspecies differentiation. International Journal of Systematic and Evolutionary Microbiology 56:501-511.
Genersch E.,2010. Honey bee pathology:current threats to honey bees and beekeeping Applied Microbiology and Biotechnology 87(1):87-97.
Gill V.,2009. "Honeybee mobs overpower hornets". BBC News. Retrieved July 5,2009.
Gilliam M.,1978. Chalkbrood status today and hopes for control. American Bee Journal 118: 468-471.
Gilliam M.,1986. Infectivity and survival of the chalkbrood pathogen, Ascosphaera apis, in colonies of honey bees, Apis mellifera. Apidologie 17:93-100.
Gilliam M., Lorenz B.J., and Buchmann S.L.,1994. Ascosphaera apis, the chalkboard pathogen of the honeybee, Apis mellifera, from larvae of a carpenter bee, Xylocopa californica arizonensis. Journal of Invertebrate Pathology 63:307-309
Gilliam M., Lorenz B.J., Prest D.B., and Camazine S.,1993. Ascosphaera apis from Apis cerana from South Korea. Journal of Invertebrate Pathology 61:111-112
Gilliam M., Lorenz J., Werner A.M., and Thorp R.W.,1997. Occurrence and distribution of Ascosphaera apis in North America:chalkbrood in feral honey bee colonies that had been in isolation on Santa Cruz Island, California for over 110 years. Apidologie 28:329-338.
Gilliam M., Taber Ⅲ S., and Rose J.B.,1978. Chalkbrood disease of honey bees, Apis mellifera L. A progress report. Apidologie 9(1):75-89.
Gilliam M., Taber S.I., Lorenz B.J., and Prest D.B.,1988. Factors affecting development of chalkbrood disease in colonies of honey bees, Apis mellifera, fed pollen contaminated with Ascosphaera apis. Journal of Invertebrate Pathology 52:314-325.
Gilliam M., and Vandenberg, J.D.,1997. Fungi. In:Morse, R., Flottum, K. (Eds.), Honey Bee Pests, Predators, and Diseases, third ed. AI Root, Ohio, OH, pp81-110.
Gilliam M.,1990. Chalkbrood disease of honey bees, Apis mellifera, caused by the fungus, Ascosphaera apis:Review of past and current research. Fifth International Colloquium on Invertebrate Pathology and Microbial Control, pp398-402.
Glinski Z., and Buczek K.,2003. Response of the Aoidea to fungal infections. Apiacta 38:183-189.
Glinski Z., and Jarosz,2000. Immune Phenomena in echinoderms. Archivum Immunologiae et Therapiae Experimentalis 48:189-193.
Glinski Z.,1982. Studies on the pathogenicity of Ascosphaera apis for honeybee brood. Part Ⅱ. Relationship between biochemical type and virulence of Ascosphaera apis. Annales Universitatis Marie curieSklodowska, DD 37:69-77.
Glinski Z.,1991. The effect of Varroa jacobsoni Oud. on the incidence and course of chalkbrood disease in Apis mellifera L. colonies. Review of Agricultural Entomology, pp 079-09747.
Gochnauer T.A., Furgala B., and Shirnamuki H.,1975. Diseases and enemies of the honey bee. In: The Hive and the Honey Bee, revised edition. Dadant Hamilton,Ill., USA, pp615-621
Gochnauer T.A., and Hughes S.J.,1976. Detection of Ascosphaera apis in honey bee brood (Hymenopte ra:Apidae) from eastern Canada. The Canadian Entomologist 108:985-988.
Gould J.L. and Gould C.G,1995. The Honey Bee. Scientific American Library. Pp19.
Govind S.,2008. Innate immunity in Drosophila:pathogens and pathways. Insect Science 15: 29-43.
Granado J.D., Kertesz-Chaloupkova K., Aebi M., and Kues U.,1997. Restriction enzyme-mediated DNA integration in Coprinus cinereus. Molecular Genomics and Genetics 256:28-36.
Haeri M., and Knox B.E.,2012. Generation of transgenic Xenopus using restriction enzyme-mediated integration. Methods in Molecular Biology 884:17-39.
Haifa B., Naima B., Naima B., Mohamed H.B.H., and Mohamed E.M.,2010. In vitro inhibitory actions of some essential oils on Ascosphaera apis, a fungus responsible for honey bee chalkbrood, Journal of Apicultural Research 49 (3):236-242
Hanif M., Pardo A.G, Gorfer M., and Raudaskoski M.,2002. T-DNA transfer and integration in the ectomycorrhizal fungus Suillus bovinus using hygromycin B as a selectable marker. Current Genetics 41:183-188.
Harbo J.,1995. Observation on hygienic behavior and resistance to chalkbrood, American Bee Journal 135:828.
Hazell B.W., Te'o V.S., Bradner J.R., Bergquist P.L., and Nevalainen K.M.,2000. Rapid transformation of high cellulase-producing mutant strains of Trichoderma reesei by microprojectile bombardment. Letters in Applied Microbiology 30:282-286.
Heath L.A.F.,1982b. Development of chalk brood in a honey bee colony:a review. Bee World 63:119-130.
Heath L.A.F.,1985. Occurrence and distribution of chalk brood disease of honeybees, Bee World 66 (1):pp9-15.
Heath L.A.F., and Gaze B.M.,1987. Carbon dioxide activation of spores of the chalkbrood fungus Ascosphaera apis. Journal of Apiculture Research 26(4):243-246.
Hedtke K., Jensen P.M., Jensen A.B., and Genersch E.,2011. Evidence for emerging parasites and pathogens influencing outbreaks of stress-related diseases like chalkbrood. Journal of Invertebrate Pathology 108:167-173.
Hepburn H.R., and Radloff S.E.,2011. Honeybees of Asia, Springer-Verlag Berlin Heidelberg. DOI 10.1007/978-3-642-16422-4.
Herrera-Estrella A., Goldman GH., and Van Montagu M.,1990. High-efficiency transformation system for the biocontrol agents, Trichoderma spp. Molecular Microbiology 4:839-843.
Hibbett D.S., Binder M.,Bischoff J.F., Blackwell M., Cannon P.F., Eriksson O.E., Huhndorf S., James T., Kirk P.M., Lucking R., Thorsten L.H., Lutzoni F., Matheny P.B., McLaughlin D.J., Powell M.J., Redhead S., Schoch C.L., Spatafora J.W., Stalpers J.A., Vilgalys R., Aime M.C.,Aptroot A., Bauer R., Begerow D., Benny G.L., Castlebury L.A., Crous P.W.,Dai Y.C., Gams W., Geiser D.M., Griffith G.W., Gueidan C., Hawksworth D.L., Hestmark G, Hosaka K., Humber R.A., Hyde K.D., Ironside J.E., Koljalg U., Kurtzman C.P., Larsson K.H., Lichtwardt R., Longcore J., Miadlikowska J., Miller A., Moncalvo J.M., Mozley-Standridge S., Oberwinkler F., Parmasto E., Reeb V, Rogers J.D., Roux C., Ryvarden L., Sampaio J.P., Schussler A., Sugiyama J., Thorn R.G, Tibell L., Untereiner W.A., Walker C., Wang Z., Weir A., Weiss M., White M.M., Winka K., Yao Y.J., and Zhang N., 2007. A higher-level phylogenetic classification of the Fungi. Mycological Research 111: 509-547.
Hitchcock J.D., and Christensen M.,1972. Occurrence of Chalkbrood(Ascosphaera Apis in Honeybees in the United States. Mycologia 64.
Holloway B., Sylvester H.A., Bourgeois L., and Rinderer T.E.,2012. Association of single nucleotide polymorphisms to resistance to chalkbrood in Apis mellifera, Journal of Apicultural Research 51(2):154-163.
Homolka L., Vyskocil P., and Pilat P.,1988. Use of protoplasts in the improvement of filamentous fungi. Appl. Microbiological Biotechnology 28:166-169.
Homolka L.,1988. Preparation, Reversion, Mutagenesis and Intra-Species Fusion of Protoplasts of Oudemansiella mucida. Fotm. Microbiology 33:298-308.
Hornitzky M.A.,2010. Honey bee diseases:Part 1. Diagnostic overview, Australia and New Zealand Standard Diagnostic Procedure, Elizabeth Macarthur Agriculture Institute, Narellan, Australia.
Hornitzky M.,2001. Literature review of chalkbrood, a fungal disease of honeybees:A report for the Rural Industries Research and Development Corporation. Publication No.01/150, Kingston, ACT, AU. pp22.
http://www.thefreedictionary.com/heterothallic:TheFreeDictionary by fablex for heterothallic definition:(accessed on January 30,2012 at 11:45pm).
Huang D., Bhairi S., and Staples R.C.,1989 A transformation procedure for Botryotinia squamosa. Current Genetics 15:411-414.
Hughes W.O.H., Eilenberg J., and Boomsma J.J.,2002. Trade-offs in group living:transmission and disease resistance in leaf-cutting ants. Proceedings of the Royal Society London B 269: 1811-1819.
Hull K.C.,2012. Restriction Enzyme Mediated Integration and Lentiviral Transgenesis of the Species Eublepharis macularius, Litoria caerulea, and Xenopus laevis. M.Sc. Thesis, North Carolina State University, Raleigh, North Carolina.
Humber R.A.,2008. Evolution of entomopathogenicity in fungi. Journal of Invertebrate Pathology 98:262-266.
Ingemar F., and Anders L.,2010. Breeding disease resistant honeybees, Swedish University of Agricultural Sciences, Uppsala, Sweden.
Invernizzi C., Rivas F., and Bettucci L.,2011. Resistance to Chalkbrood Disease in Apis mellifera L. (Hymenoptera:Apidae) Colonies with Different Hygienic Behaviour, Ecology, Behavior and Bionomics, Neotropical Entomology 40(1):28-34.
James R.R., and Buckner J.S.,2004. Lipids stimulate spore germination in the entomopathogenic ascomycete Ascosphaera aggregata. Mycozpathology 158:293-302.
James T.Y., Kauff F., Schoch C.L., Matheny P.B., Hofstetter V., Cox C.J., Celio G, Gueidan C., Fraker E., Miadlikowska J., Lumbsch H.T., Rauhut A., Reeb V., Arnold A.E., Amtoft A., Stajich J.E., Hosaka K., Sung GH., Johnson D., O'Rourke B., Crockett M., Binder M., Curtis J.M., Slot J.C., Wang Z., Wilson A.W., Schiissler A., Longcore J.E., O'Donnell K., Mozley-Standridge S., Porter D., Letcher P.M., Powell M.J., Taylor J.W., White M.M., Griffith GW., Davies D.R., Humber R.A., Morton J.B., Sugiyama J., Rossman A.Y., Rogers J.D., Pfister D.H., Hewitt D., Hansen K., Hambleton S., Shoemaker R.A., Kohlmeyer J., Volkmann-Kohlmeyer B., Spotts R.A., Serdani M., Crous P.W., Hughes K.W., Matsuura K., Langer E., Langer G, Untereiner W.A., Lucking R., Budel B., Geiser D.M., Aptroot A., Diederich P., Schmitt I., Schultz M., Yahr R., Hibbett D.S., Lutzoni F., McLaughlin D.J., Spatafora J.W., and Vilgalys R.,2006. Reconstructing the early evolution of fungi using a six-gene phylogeny. Nature 443:818-822.
Jason B., Helen E.R., and Judith K.P.,2010. Entomopathogenic fungi and insect behaviour:from unsuspecting hosts to targeted vectors, The Ecology of Fungal Entomopathogens, pp89-102.
Jensen A.B., Aronstein K., Flores J.M., Vojvodic S., Palacio M.A., and Spivak M.,2013. Standard methods for fungal brood disease research. Journal of Apicultural Research 52(1), doi: 10.3896/IBRA.1.52.1.13.
Jensen A.B., James R.R., and Eilenberg J.,2010. Long-term storage of Ascosphaera aggregata and Ascosphaera apis, pathogens of the leaf cutting bee (Megachile rotundata) and the honey bee (Apis mellifera). Journal of Invertebrate Pathology 101 (2):157-160.
Jin X., Mo M.H., Wei Z., Huang X.W., and Zhang K.Q.,2005. Transformation and mutagenesis of the nematode-trapping fungus Monacrosporium sphaeroides by restriction enzyme-mediated integration (REMI). Journal of Microbiology 43:417-423.
Kahmann R., and Basse C.,1999. REMI (restriction enzyme mediated integration) and its impact on the isolation of pathogenicity genes in fungi attacking plants. European Journal of Plant Pathology 105:221-229.
Karise R,2007. Foraging behaviour and physiology of bees:impact of insecticides. PhD-thesis, Estonian University of Life Sciences, Tartu, Estonia, pp1-123.
Kati H., Per Moestrup J., Bruun-Jensen A., and Elke G.,2011. Evidence for emerging parasites and pathogens influencing outbreaks of stress-related diseases like chalkbrood. Journal of Invertebrate Pathology 108:167-173.
Kluser S., and Peduzzi P.,2007. Global pollinator decline:a literature review. A scientific report about the current situation, recent findings and potential solution to shed light on the global pollinator crisis, UNEP/GRE), Geneva, Switzerland. pp12.
Knight C.J., Bailey A.M., and Foster GD.,2009. Agrobacterium-mediated transformation of the Plant Pathogenic Fungus Verticillium albo-atrum:Short communication. Journal of Plant Pathology 91(3):745-750.
Koenig J.P., Boush GM., and Erickson E.H.,1986. Effect of type of brood comb on chalk brood disease in honeybe colonies. Journal of Apicultural Research 25:58-62.
Lalithakumari D.,1996. Protoplasts "A biotechnological tool for plant pathological studies". Indian Phytopathology 49:199-212.
Leem Y.E., Ross I.K., and Choi H.T.,2003. Tagging and localization of a phospholipase-D-gene in Coprinellus congregatus by restriction enzyme-mediated integration and pulsed-field gel electrophoresis. FEMS Microbiological Letters 225:285-289.
Liang Q., Chen D., and Wang J.,2000. Effects of temperature, relative humidity and pH on germination of chalkbrood fungus, Ascosphaera apis spore. Chinese Journal of Applied Ecology 6:869-872.
Liu T.,1996. Varroa destructor mite as carriers of honeybee chalkbrood. American Bee Journal 136:655
Lopes F.J.F., de Queiroz M.V., Lima J.O., Silva V.A.O., and de Araujo E.F.,2008. Restriction Enzyme Improves the Efficiency of Genetic Transformations in Moniliophthora perniciosa, the Causal Agent of Witches' Broom Disease in Theobroma cacao, Brazilian Archieve of Biological Technologies 51(1):27-34.
Lorito M., Harman GE., Hayes C.K., Broadway R.M., Troncoso A., Woo S.L., and di Pietro A., 1993. Chitynolytic enzymes produced by Trichoderma harzianum:antifungal activity of purified endochitinase and chitobiase. Phytopathology 83:302-307.
Lu S., Lyngholm L., Yang G, Branson C., Yoder O.C., and Turgeon B.G,1994. Tagged mutations at the Toxl locus of Cochliobolus heterostrophus by restriction enzyme-mediated integration. Proceedings of Natural Academic Sciences, USA 91:12649-12653.
Lumbsch H.T., and Huhndorf S.M.,2007. Outline of Ascomycota. Myconet 13:1-58.
Maassen A.,1913. Weitere Mitteilungen uber der seuchenhaften Brutkrankheiten der Bienen (Further communication on the epidemic brood disease of bees). Mitteilungen aus der Kaiserlichen Biologischen Anstalt fur Land-und Forstwirtscshaft 14:48-58.
Maier F.J., and Shafer W.,1999. Mutagenesis via insertional-or restriction enzyme-mediated-integration (REMI) as a tool to tag pathogenecity related genes in plant pathogenic fungi. Biological Chemistry 380:855-864.
Martha G., Stephen Taber Ⅲ, Brenda J.L., and Dorothy B.P.,1988. Factors affecting development of chalkbrood disease in colonies of honey bees, Apis mellifera, fed pollen contaminated with Ascosphaera apis, U.S. Department of Agriculture, Agricultural Research Service, Carl Hayden Bee Research Center,2000 E. Allen Road, Tucson, Arizona 85719 USA.
Medina L.M., and Mejia E.V.,1999. The presence of Varroa jacobsoni mite and Ascosphaera apis fungi in collapsing and normal honeybee (Apis mellifera) colonies in Yucatan, Mexico. American Bee Journal 139 (10):794-796.
Michael H.,2001. Literature review of chalkbrood, a fungal disease of honeybees:A report for the Rural Industries Research and Development Corporation.
Michener C.D.,2007. The Bees of the World,2nd edition; Johns Hopkins University Press, Baltimore. pp992.
Michielse C.B., Hooykaas P.J., van den Hondel C.A., and Ram A.F.,2005. Agrobacterium-mediated transformation as a tool for functional genomics in fungi. Current Genetics 48:1-17.
Morse R.A., and Flottum K.,1997. Honey Bee Pests, Predator, & Diseases (Third edition). Root Publishing Medina, Ohio, USA, pp718.
Morse R.A., and Nowogrodzki R. (Eds.),1978. Honey Bee Pests, Predators and Diseases. (2nd Ed.). Comstock Publishing Associates, a division of Cornell University Press, Ithaca and London, ppl-10.
Murray K.D., and Aronstein K.A.,2008. Transformation of the Gram-positive honey bee pathogen, Paenibacillus larvae, by electroporation, Journal of Microbiological Methods 75:325-328.
Musallam I.W., Haddad N.J., Tawaha A.R.M., and Migdadi O.S.,2004. The Importance of Bee-Pollination in Four Genotypes of Faba Bean (Vicia faba L.). International Journal of Agriculture and Biology 6(1):9-12
Oeser B., Heidrich P.M., Muller U., Tudzynski P., and Tenberge K.B.,2002. Polygalacturonase is a pathogenicity factor in the Claviceps purpurea/rye interaction. Fungal Genetics and Biology 36:176-186.
Olmedo-Monfi V., Cortes-Penagos C., and Herrera-Estrella A.,2004. Three Decades of Fungal Transformation:Key Concepts and Applications. Methods in Molecular Biology 267: 297-313.
Palacio M.A., Pena N., Clemente G., Ruffinengo S., and Escande A.,2007. Short communication: Viability and pathogenicity of Ascosphaera apis preserved in integral rice cultures, Spanish Journal of Agricultural Research 5(4):481-486
Paul S., and Diana S.,2006. Dictionary of Microbiology and Molecular Biology, Third Edition, John Wiley & Sons Ltd. ISBN:0-470-03545-5. The Atrium, Southern Gate, Chichester, West Sussex PO19 8SQ, England.
Peberdy J.F.,1979. Fungal protoplasts:isolation, reversion, and fusion. Annual Review of Microbiology 33:21-39.
Peberdy J.F.,1991. Fungal protoplasts. In:More gene manipulations in fungi (Bennett JW, Lasure LL, eds.), Academic Press, California. pp307-318.
Peberdy J.F., and Ferenczy L. (eds.),1985. Fungal protoplasts:Applications in Biochemistry and Genetics. New York:Dekker. pp 354.
Pederson K.,1976. Chalk brood:possible methods of control, and the effect of additional heat. Birkteren 92:18-22.
Poggeler S.,2001. Mating-type genes for classical strain improvements of Ascomycetes. Applied Microbiology and Biotechnology 56:589-601.
Public Library of Science,2008. Giant Honeybees Use Shimmering 'Mexican Waves'To Repel Predatory Wasps. ScienceDaily. Retrieved March 4,2014 from www.sciencedaily.com/releases/2008/09/080909204550.htm (at 10:17 AM)
Puerto F., Flores J.M., Bustos M., Padilla F., and Campano F.,1994. Chalkbrood development in honeybee brood under controlled conditions. Apidologie 25:540-546.
Puerto F., Flores J.M., Bustos M., and Padilla F.,1995. Field and laboratory studies on chalkbrood disease of honeybees. American Bee Journal 122:29-34.
Qin X., Evans J.D., Aronstein K.A., Murray K.D., and Weinstock GM.,2006. Genome sequences of the honey bee pathogens Paenibacillus brood and Ascosphaera apis. Insect Molecular Biology.15 (5):715-718.
Rehner S.A., and Evans J.D.,2009. Microsatellite loci for the fungus Ascosphaera apis:cause of honey bee chalkbrood disease. Permanent genetic resources note, Blackwell Publishing Ltd. pp855-858.
Reynaldi F.J., Lopez A.C., Albo GN., and Alippi A.M.,2003. Genomic fingerprinting. Journal of Apiculture Research 42 (4):68-76
Riggle P.J., and Kumamoto C.A.,1998. Genetic analysis in fungi using restriction-enzyme mediated integration. Current Opinion in Microbiology 1:395-399.
Rodriguez-Tovar A.V., Ruiz-Medrano R., Herrera-Martinez A., Barrera-Figueroa B.E., Hidalgo-Lara M.E., Reyes-Marquez B.E., Cabrera-Ponce J.L., Valdes M., and Xoconostle-Cazares B.,2005. Stable genetic transformation of the ectomycorrhizal fungus Pisolithus tinctorius. Journal of Microbiological Methods 63:45-54.
Rose J.B., Christensen M., and Wilson W.T.,1984. Ascosphaera species inciting chalkbrood in North America and taxonomic key. Mycotaxon 19:41-55
Samsinakova A., Kalalova S., and Haragsin D.,1977. Effects of some antimycotics and disinfectants on the Ascosphaera apis Maassen fungus in vitro. Z. angew. Entomology 84: 225-232.
Sanchez O., Navarro R.E., and Aguirre J.,1997. Increased transformation frequency and tagging of developmental genes in Aspergillus nidulans by restriction enzyme-mediated integration (REMI). Molecular Genomics and Genetics 258:89-94.
Sankara K.R., and Prakash A.H.,1995. A simple method for the isolation of plant protoplasts. Journal of Biosciences 20:645-655.
Sato T., Yaegashi K., Ishii S., Hirano T., Kajiwara S., Shishido K., and Enei H.,1998. Transformation of the edible basidiomycete Lentinus edodes by restriction enzyme-mediated integration of plasmid DNA. Bioscience, Biotechnology and Biochemistry 62:2346-2350.
Schiestl R.H., and Petes T.D.,1991. Integration of DNA fragments by illegitimate recombination in Saccharomyces cerevisiae. Proceedings of Natural Academic Sciences, USA 88:7585-7589.
Schillberg S., Tiburzy R., and Fischer R.,2000. Transient transformation of the rust fungus Puccinia graminis f. sp. tritici. Molecular Genomics and Genetics 262:911-915.
Schmid-Hempel P.,1998. Parasites in Social Insects. Princeton University Press, Princeton, New Jersey.
Schmid-Hempel P.,2005. Evolutionary ecology of insect immune defenses. Annual Review of Entomology 50:529-551.
Schulte C., Leboulle G., Otte M., Grtinewald B., Gehne N., and Beye M.,2013. Honey bee promoter sequences for targeted gene expression, Insect Molecular Biology doi: 10.1111/imb.12031
Seong K., Hou Z., Tracy M., Kistler H.C., and Xu J.R.,2005. Random insertional mutagenesis identifies genes associated with virulence in the wheat scab fungus Fusarium graminearum. Phytopathology 95:744-750.
Shah P.A., and Pell J.K.,2003. Etnthomopathogenic fungi as biological control agents. Applied Microbiology and Biotechnology 61:413-423.
Shimanuki H., and Knox D.A.,2000. Diagnosis of Honey Bee Diseases:US Department of Agriculture, Agriculture Handbook No., AH-690:pp 61
Sikandar H., and Christos C.,2010. Isolation and fusion of protoplasts from the phytopathogenic fungus sclerotium rolfsii (sacc). Brazilian Journal of Microbiology 41:253-263
Simlesa V.,2010. Chalkbrood Disease of Honeybees, Agnote No:. Kll, Northern Territory Government, Darwin, pp3.
Skou J.P.,1972. Ascosphaerales. Friesia 10:1-24.
Skou J.P.,1988. More details in support of the class Ascosphaeromycetes. Mycotaxson ⅩⅩⅪ (1): 191-198.
Smith D.R., Villafuerte L., Otisc G., and Palmer M.R.,2000. "Biogeography of Apis cerana F. and A. nigrocincta Smith:insights from mtDNA studies" (PDF). Apidologie 31 (2):265-279.
Spiltoir C.E.,1955. Life cycle of Ascosphaera apis (Pericystis apis), American Journal of Botany 42:501-508.
Spivak M., and Downey D.L.,1998. Field assay for hygienic behavior in honey bees (Hymenoptera:Apidae). Journal of Economic Entomology 91:64-70.
Spivak M., and Gilliam M.,1993. Facultative expression of hygienic behavior in relation to disease resistance. Journal of Apiculture Research 32:147-157.
Stanley D., Yonggyun K., and Youngjin P.,2011. Cellular immunosenescence in adult male crickets, Gryllus assimilis. Archives of Insect Biochemistry and Physiology 76 (4):185-194
Strand M.R.,2008. Insect cellular immunity. Insect Science 15:1-14.
Stroschein-Stevenson S.L., Foley E., O'Farrell P.H., and Johnson A.D.,2009. Phagocytosis of Candida albicans by RNAi-treated Drosophila S2 cells. Methods in Molecular Biology 470: 347-358.
Sugahara M., and Sakamoto F.,2009. "Heat and carbon dioxide generated by honeybees jointly act to kill hornets". Naturwissenschaften 96 (9):1133-1136.
Sunagawa M., Murata H., Miyazaki Y., and Nakamura M.,2007. Transformation of the Mycorrhizal Basidiomycetes, Suillus grevillei and S. bovinus, by Particle Bombardment. Bioscince, Biotechnology and Biochemistry 71(1):47-50.
Suzuki K., Imai Y, Yamashita I., and Fukui S.,1983. In vivo ligation of linear DNA molecule to circular forms in the yeast Saccharomyces cerevisiae. Journal of Bacteriology 155:747-754.
Talkhan F.N., El-Masry H.G., Hassanein S.M., El-Beih F.M. and Abdel-Aziz M.S.,2008. Genetic improvement of dextranase Production by Penicillium funiculosum via Mutation Induction and Protoplast Fusion. International Journal of Agriculture and Biology 10(1):42-46.
Tamai Y., Miura K., and Kayama T.,1990. Electrical fusion of protoplasts between two Pleurotus species. Mokuzai Gakkaishi 36:487-490.
Tanaka A., Shiotani H., Yamamoto M., and Tsuge T.,1999. Insertional mutagenesis and cloning of the genes required for biosynthesis of the host-specific AK-toxin in the Japanese pear pathotype of Alternaria alternata. Molecular Plant Microbe Interaction 12:691-702.
Teshome G.,2006. Characterization of the causative agent of Chalkbrood disease of honeybees (Apis mellifera L.) in Ethiopia, MSc. Thesis, Addis Ababa University, School of Graduate Studies, Department of Biology, Addis Ababa, Ethiopia, pp61.
Thawley C.H.,2010. "Heat tolerance as a weapon". Davidson College. Retrieved June 1,2010.
Theantana T., and Chantawannakul P.,2008. Protease and β-N-acetylglucosaminidase of honey bee chalkbrood pathogen Ascosphaera apis, Journal of Apicultural Research 1:68-76.
Thon M.R., Nuckles E.M., and Vaillancourt L.J.,2000. Restriction Enzyme-Mediated Integration Used to Produce Pathogenicity Mutants of Colletotrichum graminicola. Molecular Plant Microbe Interation 13(12):1356-1365.
Tilburn J., Scazzocchio C., Taylor GG, Zabicky-Zissman J.H., Lockington R.A., and Davies R.W., 1983. Transformation by integration in Aspergillus nidulans. Genetics 26(2-3):205-221.
Timberlake W.E., and Marshall M.A.,1989. Genetic engineering of filamentous fungi. Science. 244:1313-1317.
Toyomatu T.M., and Mori N.D.K.,1986. Interspecific protoplast fusion between Pleurotus ostreatus and Pleurotus salmoneo-stramineus. Agricultural Biology and Chemistry 50: 223-225.
Tunlid A., Ahman J., and Oliver R.P.,1999. Transformation of the nematode-trapping fungus Arthrobotrys oligospora. FEMS Microbiological Letters 173:111-116.
Tutkun E., Maden S., Inci A., and Yilmarz B.,1993. General situation of chalkbrood disease in honeybees in Turkey. Turk Entomol Derg-TU 17 (2):65-68.
Vandenberg J.D.,1992. Bioassay of the Chalkbrood Fungus Ascosphaera aggregata on Larvae of the Alfalfa Leafcutting Bee, Megachile rotundata. Journal of Invertebrate Pathology 60: 159-163.
vanEngelsdorp D., and Marina D.M.,2010. A historical review of managed honey bee populations in Europe and the United States and the factors that may affect them. Journal of Invertebrate Pathology 103:S80-S95
Vojvodic S., Jensen A.B., Markussen B., Eilenberg J., and Boomsma J.J.,2011. Genetic Variation in Virulence among Chalkbrood Strains Infecting Honeybees, PLoS ONE 6(9):e25035.
Vojvodic S., Jensen A.B., James R.R., Boomsma J.J., and Eilenberg J.,2011. Opposing temperature-dependent mortality after infection with an obligate and facultative fungal pathogen of honeybees. Veterinary Microbiology 149:200-205.
Wang Y., Guo B., Miao Z.Q., and Tang K.,2007. Transformation of taxol-producing endophytic fungi by restriction enzyme-mediated integration (REMI), FEMS Microbiological Letters 273: 253-259.
Warwar V., Oved S., and Dickman M.B.,2000. Antisense expression of the calmodulin gene from Colletotrichum trifolii impairs prepenetration development. FEMS Microbiological Letters 191:213-219.
Wilson W.T., Nunamaker R.A., and Maki D.,1984. The occurrence of brood diseases and the absence of the Varroa mite in honeybees in Mexico. American Bee Journal 124:51-53.
Wood M.,1998. Microbes help bees battle chalkbrood. Journal of Economic Entomology 16-17.
Wubie A.J., Hu Y-Y., Li W., Huang J-X., Guo Z., Xu S-F., and Zhou T.,2013. Factors analysis in protoplast isolation and regeneration from a chalkbrood fungus, Ascosphaera apis. International Journal of Agriculture and Biology 16:89-96.
Yacobson B.A., Elad D., Rosenthal K., Kramer I., Slovecky I., and Efrat H.,1991. A recent chalkbrood outbreak in Israel:attempts at therapeutic intervention. American Bee Journal 131: 786.
Yakoby N.s Zhou R., Kobiler I., Dinoor A., and Prusky D.,2000. Development of Colletotrichum gloeosporioides Restriction Enzyme-Mediated Integration Mutants as Biocontrol Agents Against Anthracnose Disease in Avocado Fruits. The American Phytopathological Society Vol. 91, No.2,143-148. Publication no. P-2000-1218-01R
Zaghloul O.A., Mourad A.K., El Kady M.B., Nemat F.M., and Morsy M.E.,2005. Assessment of losses in honey yield due to the chalkbrood disease, with reference to the determination of its economic injury levels in Egypt. Communications in Agricultural and Applied Biological Sciences 70 (4):703-714.
Zhao K., Zhou D., Ping W., and Jingping G.,2004. Study on the Preparation and Regeneration of Protoplast from Taxol-producing Fungus Nodulisporium sylviforme; Nature and Science 2(2): 52-59.
Zhou X., Yamin W., Huifang Z., Zinan W., Juan L., Lu L., and Kexuan T.,2008, Protoplast formation, regeneration and transformation from the taxol-producing fungus Ozonium sp., African Journal of Biotechnology 7 (12):2017-2024.
Zhou X.W., Wang Z.N., Wei Y.M., Jiang K.J., Lin J., Sun X.F., Tan F., Miao Z.Q., and Tang K.X., 2006. Selection of optimal fermentation medium and enzyme system for isolation of protoplasts from endophytic fungus, Taxus mairei. Chin. Journal of Applied Environment and Biology 12(2):176-181.
Zhou X.Y., Xu S., Liu L., and Chen J.,2007. Degradation of cyanide by Trichoderma mutants constructed by restriction enzyme mediated integration (REMI). Bioresource Technology 98: 2958-2962.
Aizen M., Garibaldi L., Cunningham S., and Klein A.,2008. Long-term global trends in crop yield and production reveal no current pollination shortage but increasing pollinator dependency. Current Biology 18:1572-1575.
Aizen M., and Harder L.D.,2009. The global stock of domesticated honey bees is growing slower than agricultural demand for pollination. Current Biology 19:915-918.
Aizen M.A., Lucas A.G., Saul A.C., and Alexandra M.K.,2009. How much does agriculture depend on pollinators? Lessons from long-term trends in crop production. Annals of Botany 103: 1579-1588.
Alonso J.M, Rey J., Puerta F., Hermoso de Mendoza J., Hermoso de Mendoza M., and Flores J.M., 1993. Enzymatic equipment of Ascosphaera apis and the development of infection by this fungus in Apis mellifera. Apidologie 24:383-390.
Amaya E., and Kroll K.,2010. Production of Transgenic Xenopus laevis by Restriction Enzyme Mediated Integration and Nuclear Transplantation. Journal of Visualized Experiments 42:doi: 10.3791/2010.
Anderson D.L., Gibbs A.J., and Gibson N.L.,1998. Identification and phylogeny of spore-cyst fungi(Ascosphaera spp.) using ribosomal DNA sequences. Mycological Research 102 (5): 541-547.
Anderson D.L, and Gibson N.L.,1998. New species and isolate of spore-cysts fungi (Plectomycetes: Ascosphaerales) from Australia, Austral, Systematic Botany 11:53-72
Anderson D., Giacon H., and Gibson N.,1997. Detection and thermal destruction of the chalkbrood fungus(Ascosphaera apis) in honey. Journal of Apiculture Research 36 (3-4):163-168.
Aronstein K.A, Murray K.D, and Saldivar E.,2010. Transcriptional responses in Honey Bee larvae infected with chalkbrood fungus. BMC Genomics 11:pp391.
Aronstein K.A, and Murray K.D.,2010. Chalkbrood disease in honey bees, Journal of Invertebrate Pathology 103:S20-S29.
Aupinel P., Fortini D., Dufour H., Tasei J.N., Michaud B., Odoux J.F., and Pham-Delegue M.H., 2005. Improvement of artificial feeding in a standard in vitro method for rearing Apis mellifera larvae. Bulletin of Insectology 58:107-111.
Aupinel P., Fortini D., Michaud B., Marolleau F., Tasel J.N., and Odoux J.F.,2007a. Toxicity of dimethoate and fenoxycarb to honey bee brood(Apis mellifera), using a new in vitro standardized feeding method. Pest Management Science 63:1090-1094.
Aupinel P., Fortini D., Michaud B., Medrzycki P., Padovani E., Przygoda D., Maus C., Charriere J.D., Kilchenmann V., Riessberger-Galle U., Vollmann J.J., Jeker L., Janke M., Odoux J.F., and Tasei J.N.,2009. Honey bee brood ring-test:method for testing pesticide toxicity on honey bee brood in laboratory conditions. Julius-Kuhn-Archiv 423:96-102.
Aupinel P., Medrzycki P., Fortini D., Michaud B., Tasei J.N., and Odoux J.F.,2007b. A new larval in vitro rearing method to test effects of pesticides on honey bee brood. Redia 90:91-94.
Bailey L.,1963. Infectious Diseases of the Honeybee, Land Books Ltd. London, pp176.
Bailey L.,1967. The effect of temperature on the pathogenicity of the fungus Ascosphaera apis, for brood of the honey bee, Apis mellifera. In Proceedings of the International Cooloquium on Insect Pathology and Microbial Control (P.A. vab der Laan, Ed. pp162-167.
Bailey L., and Ball B.V.,1991. Honey Bee Pathology. Academic Press, London, pp53-63; pp154-158
Bailey L., and Fernando, E.F.W.,1972. Effects of sacbrood virus on adult honey bees. Annals of Applied Biology 72:27-35.
Bakhiet N., and Stahly D.P.,1985. Ultrastructure of Sporulating Bacillus larvae in a Broth Medium. Applied and Environmental Microbiology 50(3):690-692.
Balhareth H.M., Alqarni, A.S. and Owayss, A.A.,2012. Comparison of hygienic and grooming behaviors of indigenous and exotic honeybee, Apis mellifera races in central Saudi Arabia. International Journal of Agriculture and Biology 14:1005-1008.
Bamford S.,1987. Studies on the infection of honeybee larvae with Ascosphaera apis. PhD Thesis, Plymouth Polytechnic, United Kingdom.
Bamford S., and Heath L.A.F.,1989. The effects of temperature and pH on the germination of spores of the Chalkbrood fungus,A scosphaera apis. Journal of Apicultural Research,28: 36-40.
Barnett E.A., Charlton A.J., and Fletcher M.R.,2007. Incidents of bee poisoning with pesticides in the United Kingdom 1994-2003, Pest Management Sciences 63:1051-1057
Benedek P.,1985. Economic importance of honeybee pollination of crops at the national level in Hungary, Proceedings of the 29th international beekeeping congress, Budapest, Hungary, Apimondia, pp286-289.
Bills S.N., Richter, D.L., and Podila, GK.,1995. Genetic transformation of the ectomycorrhizal fungus Paxillus involutus by particle bombardment. Mycological Research 99:557-561
Bissett J., Duke G, and Goettel M.,1996. Ascosphaera acerosa species nov. Isolated from the alfaalfa leafcutting bee, with a key to the species of Ascosphaera, Mycologia 88 (5):797-803
Bissett J.,1988. Contribution towards a monograph of the genus Ascosphaera. Canadian Journal of Botany 66:2541-2560.
Blochlinger K., and Diggelmann H.,1984. Hygromycin B Phosphotransferase as a Selectable Marker for DNA Transfer Experiments with Higher Eucaryotic Cells. Molecular Cell Biology 4(12):2929-2931.
Borum A.E., and Ulgen M.,2008. Chalkbrood(Ascosphaera apis) infection and fungal agents of honey bees in North-West Turkey. Journal of Apicultural Research 47(2):170-171
Bruun-Jensen A., Bo V.P., and Jorgen E.,2009. Differential susceptibility across honey bee colonies in larval chalkbrood resistance, Apidologie 40:524-534.
Camazine S.,1989. Beekeeping in South Korea. Bee World 70:66-74.
Campano F., Flores J.M., Puerta F., Ruiz J.A., and Ruz J.M.,1999. Fungal diseases of the honeybee (Apis mellifera L.). In:Colin M.E. (ed.), Ball B.V. (ed.), Kilani M. (ed.). Bee disease diagnosis. Zaragoza:CIHEAM, (Options Mediterraneennes:Serie B. Etudes et Recherches; n.25), pp61-68.
Carrera P., Sommaruga A., and Vailati G.,1987. The development of Ascosphaera apis within larvae of Apis mellifera ligustica. Journal of Apiculture Research 26(1):59-63.
Case M.E., Schweizer M., Kushner S.R., Giles N.H.,1979. Efficient transformation of Neurospora crassa by utilizing hybrid plasmid DNA. Proc. Natural Academic Seinces, USA.76: 5259-5263.
Chen R., and Jeffrey J.M.,1993. Production of Fungal Protoplasts from selected wood-degrading fungi. Oregon State University, Wood and Fiber Science 25 (1):61-65.
Chen Y.P. and Siede R.,2007. Honey bee viruses. Advances in Virus Research 70:33-80.
Chorbinski P., and Rypula K.,2003. Studies on the morphology of strains Ascosphaera apis isolated from chalkbrood disease of the honey bees. Veterinary Medicine 6 (2):1-12.
Chorbinski P.,2004. Identification of Ascosphaera apis strains using ribosomal DNA sequences. Medycyna Weterynaryjna 60(2):190-192.
Chung K.R., Ehrenshaft M., Wetzel D.K., and Daub M.E.,2003. Cercosporin-deficient mutants by plasmid tagging in the asexual fungus Cercospora nicotianae. Molecular Genetics and Genomics270:103-113.
Cocking E.C.,1960. A method for the isolation of plant protoplasts and vacuoles; Nature (London) 187:927-929.
Cocking E.C.,1979. All sorts of protoplasts; Nature,281:180-181. Davis BD,1985. Factors affecting protoplast isolation. In:Fungal protoplasts:Application in biochemistry and genetics (Peberdy JF, Ferenczy L eds.), Marcel Dekker, New York, pp45-72.
Cordoni G.,2011. Epidemiology and taxonomy of honey bee viruses in England and Wales. PhD thesis. University of Surrey.
Cordoni G., and Spagnuolo M.L.,2007. Development of an experimental GIS for bee-keeping in the Marche Region, Italy. Veterinaria Italiana 43(3):431-436.
Cornman R.S., Bennett A.K., Murray K.D., Evans J.D., Elsik C.G., and Aronstein K.,2012. Transcriptome analysis of the honey bee fungal pathogen, Ascosphaera apis:implications for host pathogenesis. BMC Genomics 13:pp285.
Cove D.J.,1979. The uses of isolated protoplasts in plant genetics. Heredity 43 (3):295-314.
Crailsheim K., Brodschneider R., Aupinel P., Behrens D., Genersch E., Vollmann J., and Riessberger-Galle U.,2013. Standard methods for artificial rearing of Apis mellifera larvae. Journal of Apiculture Research 52(1):1-15.
Crane E.,1991. Apis species of tropical Asia as pollinators and some rearing methods for them. Acta Horticulture 288:29-48.
Crane E.,1999. The World History of Beekeeping and Honey Hunting. New York:Routledge.
Cremer S., Jiirgen H., Alexandra S., Daniel J.C.K., and Line V.U.,2010. Rapid anti-pathogen response in ant societies relies on high genetic diversity. Proceedings of the Royal Society B: Biological Sciences 277:2821-2828
Cummings W.J., Celerin M., Crodian J., Brunick L.K., and Zolan M.E.,1999. Insertional mutageneses in Coprinus cinereus:use of a dominant selectable marker to generate tagged, sporulation-defective mutants. Current Genetics 36:371-382.
Cundliffe E.,1968. Preparation and Some Properties of Active Protoplasts of Bacillus megaterium, Journal of General Microbiology 53:425-430.
Davey M.R., Anthony P., Power J.B., and Lowe K.C.,2005. Plant protoplasts:status and biotechnological Perspectives. Biotechnological Advances 23:131-171.
Davis C., and Ward W.,2003. Control of chalkbrood disease with natural products:a report for the RIRDC. Publication No.03/107, Kingston, ACT, AU, ppl-23.
Deodikar G.B., Ghatge A.L., Phadke R.P., Mahindre D.B., Kshirsagar K.K., Muvel S., and Thakar S.S.,1977. Nesting behaviour of Indian honeybees. Ⅲ. Nesting behaviour of Apis dorsata. Indian Bee Journal 39:1-12.
Desalegn B.,2006. The Occurrence of Chalk Brood (Ascosphaera apis):A Honeybee (A. mellifera L.) Disease in West Shoa, Ethiopia, Ethiopian Journal of Animal Production 6(1):1-8.
Desneux N., Decourtye A., and Delpuech J.M.,2007. The sub lethal effects of pesticides on beneficial arthropods. Annual Review of Entomology 52:81-106.
Eguchi F., Renato G.R., Tomoaki I., and Miyato H.,1998. Regeneration of protoplasts from hyphal strands of Volvariella volvacea Journal of Wood Science 44(5):401-407.
Eguchi F., Tashiro M., Suzuki T., and Higaki M.,1990. Preparation and regeneration of protoplasts of edible mushrooms. Mokuzai Gakkaishi 36:232-240.
Engel M.S.,1999. "The taxonomy of recent and fossil honey bees (Hymenoptera:Apidae:Apis)". Journal of Hymenoptera Research 8:165-196.
Engel M.S., Hinojosa-Diaz I.A., and Rasnitsyn A.P.,2009. "A honey bee from the Miocene of Nevada and the biogeography of Apis (Hymenoptera:Apidae:Apini)". Proceedings of the California Academy of Sciences 60 (3):23-38.
European Synchrotron Radiation Facility,2012. First ever record of insect pollination from 100 million years ago. ScienceDaily. Retrieved October 8,2013, from http://www.sciencedaily.com/releases/2012/05/120514153113.htm.
Evans J.D., Aronstein K., Chen Y.P., Hetru C., Imler J.L., Jiang H., Kanost M., Thompson G.J., Zou Z., and Hultmark D.,2006. Immune pathways and defence mechanisms in honey bees Apis mellifera. Insect Molecular Biology 15(5):645-656.
Evans J.D., and Spivak M.,2010. Socialized medicine:Individual and communal disease barriers in honey bees. Journal of Invertebrate Pathology 103:S62-S72.
Feng F.,2000. Infectious diseases in honeybee and the control methods. Journal of bee (7):pp19
Fincham J.R.S.,1989. Transformation in fungi. Microbiological Review 53:148-170.
Flores J.M., Gutierrez I., and Espejo R.,2005b. The role of pollen in chalkbrood disease in Apis mellifera:transmission and predisposing conditions. Mycologia 97:1171-1176.
Flores J.M., Gutierrez I., and Puerta F.,2004a. A comparison of methods to experimentally induce chalkbrood disease in honey bees. Spanish Journal of Agricultural Research 2 (1):79-83.
Flores J.M., Gutierrez I., and Puerta F.,2004b. Oxytetracycline as a predisposing condition for chalkbrood in honeybee. Veterinary Microbiology 103:195-199.
Flores J., Spivak M., and Gutierrez I.,2005a. Spores of Ascosphaera apis contained in wax foundation can infect honeybee brood. Veterinary Microbiology 108:141-144.
Flores J.M., Ruiz J.A., Ruz J.M., Puerta F., Bustos M., Padilla F., and Campano F.,1996. Effect of temperature and humidity of sealed brood on chalkbrood development under controlled conditions. Apidologie 27:185-192.
Flowers J.L., and Vaillancourt L.J.,2005. Parameters affecting the efficiency of Agrobacterium tumefaciensmediated transformation of Colletotrichum graminicola. Current Genetics 48: 380-388.
Fries I., and Camazine S.,2001. Implications of horizontal and vertical pathogen transmission for honey bee epidemiology. Apidologie 32(3):199-214.
Fries I., and Lindstrom A.,2010. Breeding disease resistant honeybees, Swedish University of Agricultural Sciences, Uppsala, Sweden. pp21.
Fujimoto D., Shi Y., Christian D., Mantaguihan J.B., and Leung H.,2002 Tagging quantitative loci controlling pathogenicity in Magnaporthe grisea by insertional mutagenesis. Physiological and Molecular Plant Pathology 61:77-88.
Fungaro M.H.P., Rech E., Muhlen GS., Vainstein M.H., Pascon R.C., de Queiroz M.V., Pizzirani-Kleiner A.A., and de Azevedo J.L.,1995. Transformation of Aspergillus nidulans by microprojectile bombardment on intact conidia. FEMS Microbiological Letters 125:293-298.
Gallai N., Salles J.M., Settele J., and Vaissie're B.E.,2009. Economic valuation of the vulnerability of world agriculture confronted with pollinator decline. Ecological Economics 68:810-821.
Gao C., Xue Y., and Ma Y,2011. Protoplast Transformation of Recalcitrant Alkaliphilic Bacillus sp. with Methylated Plasmid DNA and a Developed Hard Agar Regeneration Medium; PLoS ONE 6(11):e28148. doi:10.1371/journal.pone.0028148
Genersch E., Ashiralieva A., and Fries I.,2005. Strain- and genotype-specific differences in virulence of Paenibacillus larvae subsp. larvae, a bacterial pathogen causing American foulbrood disease in honey bees. Applied Environmental Microbiology, pp7551-7555. http://dx.doi.org/10.1128/AEM.71.11.7551-7555.2005.
Genersch E., Forsgren E., Pentikainen J., Ashiralieva A., Rauch S., Kilwinski J., and Fries I.,2006. Reclassification of Paenibacillus larvae subsp. pulvifaciens and Paenibacillus larvae subsp. larvae as Paenibacillus larvae without subspecies differentiation. International Journal of Systematic and Evolutionary Microbiology 56:501-511.
Genersch E.,2010. Honey bee pathology:current threats to honey bees and beekeeping Applied Microbiology and Biotechnology 87(1):87-97.
Gill V.,2009. "Honeybee mobs overpower hornets". BBC News. Retrieved July 5,2009.
Gilliam M.,1978. Chalkbrood status today and hopes for control. American Bee Journal 118: 468-471.
Gilliam M.,1986. Infectivity and survival of the chalkbrood pathogen, Ascosphaera apis, in colonies of honey bees, Apis mellifera. Apidologie 17:93-100.
Gilliam M., Lorenz B.J., and Buchmann S.L.,1994. Ascosphaera apis, the chalkboard pathogen of the honeybee, Apis mellifera, from larvae of a carpenter bee, Xylocopa californica arizonensis. Journal of Invertebrate Pathology 63:307-309
Gilliam M., Lorenz B.J., Prest D.B., and Camazine S.,1993. Ascosphaera apis from Apis cerana from South Korea. Journal of Invertebrate Pathology 61:111-112
Gilliam M., Lorenz J., Werner A.M., and Thorp R.W.,1997. Occurrence and distribution of Ascosphaera apis in North America:chalkbrood in feral honey bee colonies that had been in isolation on Santa Cruz Island, California for over 110 years. Apidologie 28:329-338.
Gilliam M., Taber Ⅲ S., and Rose J.B.,1978. Chalkbrood disease of honey bees, Apis mellifera L. A progress report. Apidologie 9(1):75-89.
Gilliam M., Taber S.I., Lorenz B.J., and Prest D.B.,1988. Factors affecting development of chalkbrood disease in colonies of honey bees, Apis mellifera, fed pollen contaminated with Ascosphaera apis. Journal of Invertebrate Pathology 52:314-325.
Gilliam M., and Vandenberg, J.D.,1997. Fungi. In:Morse, R., Flottum, K. (Eds.), Honey Bee Pests, Predators, and Diseases, third ed. AI Root, Ohio, OH, pp81-110.
Gilliam M.,1990. Chalkbrood disease of honey bees, Apis mellifera, caused by the fungus, Ascosphaera apis:Review of past and current research. Fifth International Colloquium on Invertebrate Pathology and Microbial Control, pp398-402.
Glinski Z., and Buczek K.,2003. Response of the Aoidea to fungal infections. Apiacta 38:183-189.
Glinski Z., and Jarosz,2000. Immune Phenomena in echinoderms. Archivum Immunologiae et Therapiae Experimentalis 48:189-193.
Glinski Z.,1982. Studies on the pathogenicity of Ascosphaera apis for honeybee brood. Part Ⅱ. Relationship between biochemical type and virulence of Ascosphaera apis. Annales Universitatis Marie curieSklodowska, DD 37:69-77.
Glinski Z.,1991. The effect of Varroa jacobsoni Oud. on the incidence and course of chalkbrood disease in Apis mellifera L. colonies. Review of Agricultural Entomology, pp 079-09747.
Gochnauer T.A., Furgala B., and Shirnamuki H.,1975. Diseases and enemies of the honey bee. In: The Hive and the Honey Bee, revised edition. Dadant Hamilton,Ill., USA, pp615-621
Gochnauer T.A., and Hughes S.J.,1976. Detection of Ascosphaera apis in honey bee brood (Hymenopte ra:Apidae) from eastern Canada. The Canadian Entomologist 108:985-988.
Gould J.L. and Gould C.G,1995. The Honey Bee. Scientific American Library. Pp19.
Govind S.,2008. Innate immunity in Drosophila:pathogens and pathways. Insect Science 15: 29-43.
Granado J.D., Kertesz-Chaloupkova K., Aebi M., and Kues U.,1997. Restriction enzyme-mediated DNA integration in Coprinus cinereus. Molecular Genomics and Genetics 256:28-36.
Haeri M., and Knox B.E.,2012. Generation of transgenic Xenopus using restriction enzyme-mediated integration. Methods in Molecular Biology 884:17-39.
Haifa B., Naima B., Naima B., Mohamed H.B.H., and Mohamed E.M.,2010. In vitro inhibitory actions of some essential oils on Ascosphaera apis, a fungus responsible for honey bee chalkbrood, Journal of Apicultural Research 49 (3):236-242
Hanif M., Pardo A.G, Gorfer M., and Raudaskoski M.,2002. T-DNA transfer and integration in the ectomycorrhizal fungus Suillus bovinus using hygromycin B as a selectable marker. Current Genetics 41:183-188.
Harbo J.,1995. Observation on hygienic behavior and resistance to chalkbrood, American Bee Journal 135:828.
Hazell B.W., Te'o V.S., Bradner J.R., Bergquist P.L., and Nevalainen K.M.,2000. Rapid transformation of high cellulase-producing mutant strains of Trichoderma reesei by microprojectile bombardment. Letters in Applied Microbiology 30:282-286.
Heath L.A.F.,1982b. Development of chalk brood in a honey bee colony:a review. Bee World 63:119-130.
Heath L.A.F.,1985. Occurrence and distribution of chalk brood disease of honeybees, Bee World 66 (1):pp9-15.
Heath L.A.F., and Gaze B.M.,1987. Carbon dioxide activation of spores of the chalkbrood fungus Ascosphaera apis. Journal of Apiculture Research 26(4):243-246.
Hedtke K., Jensen P.M., Jensen A.B., and Genersch E.,2011. Evidence for emerging parasites and pathogens influencing outbreaks of stress-related diseases like chalkbrood. Journal of Invertebrate Pathology 108:167-173.
Hepburn H.R., and Radloff S.E.,2011. Honeybees of Asia, Springer-Verlag Berlin Heidelberg. DOI 10.1007/978-3-642-16422-4.
Herrera-Estrella A., Goldman GH., and Van Montagu M.,1990. High-efficiency transformation system for the biocontrol agents, Trichoderma spp. Molecular Microbiology 4:839-843.
Hibbett D.S., Binder M.,Bischoff J.F., Blackwell M., Cannon P.F., Eriksson O.E., Huhndorf S., James T., Kirk P.M., Lucking R., Thorsten L.H., Lutzoni F., Matheny P.B., McLaughlin D.J., Powell M.J., Redhead S., Schoch C.L., Spatafora J.W., Stalpers J.A., Vilgalys R., Aime M.C.,Aptroot A., Bauer R., Begerow D., Benny G.L., Castlebury L.A., Crous P.W.,Dai Y.C., Gams W., Geiser D.M., Griffith G.W., Gueidan C., Hawksworth D.L., Hestmark G, Hosaka K., Humber R.A., Hyde K.D., Ironside J.E., Koljalg U., Kurtzman C.P., Larsson K.H., Lichtwardt R., Longcore J., Miadlikowska J., Miller A., Moncalvo J.M., Mozley-Standridge S., Oberwinkler F., Parmasto E., Reeb V, Rogers J.D., Roux C., Ryvarden L., Sampaio J.P., Schussler A., Sugiyama J., Thorn R.G, Tibell L., Untereiner W.A., Walker C., Wang Z., Weir A., Weiss M., White M.M., Winka K., Yao Y.J., and Zhang N., 2007. A higher-level phylogenetic classification of the Fungi. Mycological Research 111: 509-547.
Hitchcock J.D., and Christensen M.,1972. Occurrence of Chalkbrood(Ascosphaera Apis in Honeybees in the United States. Mycologia 64.
Holloway B., Sylvester H.A., Bourgeois L., and Rinderer T.E.,2012. Association of single nucleotide polymorphisms to resistance to chalkbrood in Apis mellifera, Journal of Apicultural Research 51(2):154-163.
Homolka L., Vyskocil P., and Pilat P.,1988. Use of protoplasts in the improvement of filamentous fungi. Appl. Microbiological Biotechnology 28:166-169.
Homolka L.,1988. Preparation, Reversion, Mutagenesis and Intra-Species Fusion of Protoplasts of Oudemansiella mucida. Fotm. Microbiology 33:298-308.
Hornitzky M.A.,2010. Honey bee diseases:Part 1. Diagnostic overview, Australia and New Zealand Standard Diagnostic Procedure, Elizabeth Macarthur Agriculture Institute, Narellan, Australia.
Hornitzky M.,2001. Literature review of chalkbrood, a fungal disease of honeybees:A report for the Rural Industries Research and Development Corporation. Publication No.01/150, Kingston, ACT, AU. pp22.
http://www.thefreedictionary.com/heterothallic:TheFreeDictionary by fablex for heterothallic definition:(accessed on January 30,2012 at 11:45pm).
Huang D., Bhairi S., and Staples R.C.,1989 A transformation procedure for Botryotinia squamosa. Current Genetics 15:411-414.
Hughes W.O.H., Eilenberg J., and Boomsma J.J.,2002. Trade-offs in group living:transmission and disease resistance in leaf-cutting ants. Proceedings of the Royal Society London B 269: 1811-1819.
Hull K.C.,2012. Restriction Enzyme Mediated Integration and Lentiviral Transgenesis of the Species Eublepharis macularius, Litoria caerulea, and Xenopus laevis. M.Sc. Thesis, North Carolina State University, Raleigh, North Carolina.
Humber R.A.,2008. Evolution of entomopathogenicity in fungi. Journal of Invertebrate Pathology 98:262-266.
Ingemar F., and Anders L.,2010. Breeding disease resistant honeybees, Swedish University of Agricultural Sciences, Uppsala, Sweden.
Invernizzi C., Rivas F., and Bettucci L.,2011. Resistance to Chalkbrood Disease in Apis mellifera L. (Hymenoptera:Apidae) Colonies with Different Hygienic Behaviour, Ecology, Behavior and Bionomics, Neotropical Entomology 40(1):28-34.
James R.R., and Buckner J.S.,2004. Lipids stimulate spore germination in the entomopathogenic ascomycete Ascosphaera aggregata. Mycozpathology 158:293-302.
James T.Y., Kauff F., Schoch C.L., Matheny P.B., Hofstetter V., Cox C.J., Celio G, Gueidan C., Fraker E., Miadlikowska J., Lumbsch H.T., Rauhut A., Reeb V., Arnold A.E., Amtoft A., Stajich J.E., Hosaka K., Sung GH., Johnson D., O'Rourke B., Crockett M., Binder M., Curtis J.M., Slot J.C., Wang Z., Wilson A.W., Schiissler A., Longcore J.E., O'Donnell K., Mozley-Standridge S., Porter D., Letcher P.M., Powell M.J., Taylor J.W., White M.M., Griffith GW., Davies D.R., Humber R.A., Morton J.B., Sugiyama J., Rossman A.Y., Rogers J.D., Pfister D.H., Hewitt D., Hansen K., Hambleton S., Shoemaker R.A., Kohlmeyer J., Volkmann-Kohlmeyer B., Spotts R.A., Serdani M., Crous P.W., Hughes K.W., Matsuura K., Langer E., Langer G, Untereiner W.A., Lucking R., Budel B., Geiser D.M., Aptroot A., Diederich P., Schmitt I., Schultz M., Yahr R., Hibbett D.S., Lutzoni F., McLaughlin D.J., Spatafora J.W., and Vilgalys R.,2006. Reconstructing the early evolution of fungi using a six-gene phylogeny. Nature 443:818-822.
Jason B., Helen E.R., and Judith K.P.,2010. Entomopathogenic fungi and insect behaviour:from unsuspecting hosts to targeted vectors, The Ecology of Fungal Entomopathogens, pp89-102.
Jensen A.B., Aronstein K., Flores J.M., Vojvodic S., Palacio M.A., and Spivak M.,2013. Standard methods for fungal brood disease research. Journal of Apicultural Research 52(1), doi: 10.3896/IBRA.1.52.1.13.
Jensen A.B., James R.R., and Eilenberg J.,2010. Long-term storage of Ascosphaera aggregata and Ascosphaera apis, pathogens of the leaf cutting bee (Megachile rotundata) and the honey bee (Apis mellifera). Journal of Invertebrate Pathology 101 (2):157-160.
Jin X., Mo M.H., Wei Z., Huang X.W., and Zhang K.Q.,2005. Transformation and mutagenesis of the nematode-trapping fungus Monacrosporium sphaeroides by restriction enzyme-mediated integration (REMI). Journal of Microbiology 43:417-423.
Kahmann R., and Basse C.,1999. REMI (restriction enzyme mediated integration) and its impact on the isolation of pathogenicity genes in fungi attacking plants. European Journal of Plant Pathology 105:221-229.
Karise R,2007. Foraging behaviour and physiology of bees:impact of insecticides. PhD-thesis, Estonian University of Life Sciences, Tartu, Estonia, pp1-123.
Kati H., Per Moestrup J., Bruun-Jensen A., and Elke G.,2011. Evidence for emerging parasites and pathogens influencing outbreaks of stress-related diseases like chalkbrood. Journal of Invertebrate Pathology 108:167-173.
Kluser S., and Peduzzi P.,2007. Global pollinator decline:a literature review. A scientific report about the current situation, recent findings and potential solution to shed light on the global pollinator crisis, UNEP/GRE), Geneva, Switzerland. pp12.
Knight C.J., Bailey A.M., and Foster GD.,2009. Agrobacterium-mediated transformation of the Plant Pathogenic Fungus Verticillium albo-atrum:Short communication. Journal of Plant Pathology 91(3):745-750.
Koenig J.P., Boush GM., and Erickson E.H.,1986. Effect of type of brood comb on chalk brood disease in honeybe colonies. Journal of Apicultural Research 25:58-62.
Lalithakumari D.,1996. Protoplasts "A biotechnological tool for plant pathological studies". Indian Phytopathology 49:199-212.
Leem Y.E., Ross I.K., and Choi H.T.,2003. Tagging and localization of a phospholipase-D-gene in Coprinellus congregatus by restriction enzyme-mediated integration and pulsed-field gel electrophoresis. FEMS Microbiological Letters 225:285-289.
Liang Q., Chen D., and Wang J.,2000. Effects of temperature, relative humidity and pH on germination of chalkbrood fungus, Ascosphaera apis spore. Chinese Journal of Applied Ecology 6:869-872.
Liu T.,1996. Varroa destructor mite as carriers of honeybee chalkbrood. American Bee Journal 136:655
Lopes F.J.F., de Queiroz M.V., Lima J.O., Silva V.A.O., and de Araujo E.F.,2008. Restriction Enzyme Improves the Efficiency of Genetic Transformations in Moniliophthora perniciosa, the Causal Agent of Witches' Broom Disease in Theobroma cacao, Brazilian Archieve of Biological Technologies 51(1):27-34.
Lorito M., Harman GE., Hayes C.K., Broadway R.M., Troncoso A., Woo S.L., and di Pietro A., 1993. Chitynolytic enzymes produced by Trichoderma harzianum:antifungal activity of purified endochitinase and chitobiase. Phytopathology 83:302-307.
Lu S., Lyngholm L., Yang G, Branson C., Yoder O.C., and Turgeon B.G,1994. Tagged mutations at the Toxl locus of Cochliobolus heterostrophus by restriction enzyme-mediated integration. Proceedings of Natural Academic Sciences, USA 91:12649-12653.
Lumbsch H.T., and Huhndorf S.M.,2007. Outline of Ascomycota. Myconet 13:1-58.
Maassen A.,1913. Weitere Mitteilungen uber der seuchenhaften Brutkrankheiten der Bienen (Further communication on the epidemic brood disease of bees). Mitteilungen aus der Kaiserlichen Biologischen Anstalt fur Land-und Forstwirtscshaft 14:48-58.
Maier F.J., and Shafer W.,1999. Mutagenesis via insertional-or restriction enzyme-mediated-integration (REMI) as a tool to tag pathogenecity related genes in plant pathogenic fungi. Biological Chemistry 380:855-864.
Martha G., Stephen Taber Ⅲ, Brenda J.L., and Dorothy B.P.,1988. Factors affecting development of chalkbrood disease in colonies of honey bees, Apis mellifera, fed pollen contaminated with Ascosphaera apis, U.S. Department of Agriculture, Agricultural Research Service, Carl Hayden Bee Research Center,2000 E. Allen Road, Tucson, Arizona 85719 USA.
Medina L.M., and Mejia E.V.,1999. The presence of Varroa jacobsoni mite and Ascosphaera apis fungi in collapsing and normal honeybee (Apis mellifera) colonies in Yucatan, Mexico. American Bee Journal 139 (10):794-796.
Michael H.,2001. Literature review of chalkbrood, a fungal disease of honeybees:A report for the Rural Industries Research and Development Corporation.
Michener C.D.,2007. The Bees of the World,2nd edition; Johns Hopkins University Press, Baltimore. pp992.
Michielse C.B., Hooykaas P.J., van den Hondel C.A., and Ram A.F.,2005. Agrobacterium-mediated transformation as a tool for functional genomics in fungi. Current Genetics 48:1-17.
Morse R.A., and Flottum K.,1997. Honey Bee Pests, Predator, & Diseases (Third edition). Root Publishing Medina, Ohio, USA, pp718.
Morse R.A., and Nowogrodzki R. (Eds.),1978. Honey Bee Pests, Predators and Diseases. (2nd Ed.). Comstock Publishing Associates, a division of Cornell University Press, Ithaca and London, ppl-10.
Murray K.D., and Aronstein K.A.,2008. Transformation of the Gram-positive honey bee pathogen, Paenibacillus larvae, by electroporation, Journal of Microbiological Methods 75:325-328.
Musallam I.W., Haddad N.J., Tawaha A.R.M., and Migdadi O.S.,2004. The Importance of Bee-Pollination in Four Genotypes of Faba Bean (Vicia faba L.). International Journal of Agriculture and Biology 6(1):9-12
Oeser B., Heidrich P.M., Muller U., Tudzynski P., and Tenberge K.B.,2002. Polygalacturonase is a pathogenicity factor in the Claviceps purpurea/rye interaction. Fungal Genetics and Biology 36:176-186.
Olmedo-Monfi V., Cortes-Penagos C., and Herrera-Estrella A.,2004. Three Decades of Fungal Transformation:Key Concepts and Applications. Methods in Molecular Biology 267: 297-313.
Palacio M.A., Pena N., Clemente G., Ruffinengo S., and Escande A.,2007. Short communication: Viability and pathogenicity of Ascosphaera apis preserved in integral rice cultures, Spanish Journal of Agricultural Research 5(4):481-486
Paul S., and Diana S.,2006. Dictionary of Microbiology and Molecular Biology, Third Edition, John Wiley & Sons Ltd. ISBN:0-470-03545-5. The Atrium, Southern Gate, Chichester, West Sussex PO19 8SQ, England.
Peberdy J.F.,1979. Fungal protoplasts:isolation, reversion, and fusion. Annual Review of Microbiology 33:21-39.
Peberdy J.F.,1991. Fungal protoplasts. In:More gene manipulations in fungi (Bennett JW, Lasure LL, eds.), Academic Press, California. pp307-318.
Peberdy J.F., and Ferenczy L. (eds.),1985. Fungal protoplasts:Applications in Biochemistry and Genetics. New York:Dekker. pp 354.
Pederson K.,1976. Chalk brood:possible methods of control, and the effect of additional heat. Birkteren 92:18-22.
Poggeler S.,2001. Mating-type genes for classical strain improvements of Ascomycetes. Applied Microbiology and Biotechnology 56:589-601.
Public Library of Science,2008. Giant Honeybees Use Shimmering 'Mexican Waves'To Repel Predatory Wasps. ScienceDaily. Retrieved March 4,2014 from www.sciencedaily.com/releases/2008/09/080909204550.htm (at 10:17 AM)
Puerto F., Flores J.M., Bustos M., Padilla F., and Campano F.,1994. Chalkbrood development in honeybee brood under controlled conditions. Apidologie 25:540-546.
Puerto F., Flores J.M., Bustos M., and Padilla F.,1995. Field and laboratory studies on chalkbrood disease of honeybees. American Bee Journal 122:29-34.
Qin X., Evans J.D., Aronstein K.A., Murray K.D., and Weinstock GM.,2006. Genome sequences of the honey bee pathogens Paenibacillus brood and Ascosphaera apis. Insect Molecular Biology.15 (5):715-718.
Rehner S.A., and Evans J.D.,2009. Microsatellite loci for the fungus Ascosphaera apis:cause of honey bee chalkbrood disease. Permanent genetic resources note, Blackwell Publishing Ltd. pp855-858.
Reynaldi F.J., Lopez A.C., Albo GN., and Alippi A.M.,2003. Genomic fingerprinting. Journal of Apiculture Research 42 (4):68-76
Riggle P.J., and Kumamoto C.A.,1998. Genetic analysis in fungi using restriction-enzyme mediated integration. Current Opinion in Microbiology 1:395-399.
Rodriguez-Tovar A.V., Ruiz-Medrano R., Herrera-Martinez A., Barrera-Figueroa B.E., Hidalgo-Lara M.E., Reyes-Marquez B.E., Cabrera-Ponce J.L., Valdes M., and Xoconostle-Cazares B.,2005. Stable genetic transformation of the ectomycorrhizal fungus Pisolithus tinctorius. Journal of Microbiological Methods 63:45-54.
Rose J.B., Christensen M., and Wilson W.T.,1984. Ascosphaera species inciting chalkbrood in North America and taxonomic key. Mycotaxon 19:41-55
Samsinakova A., Kalalova S., and Haragsin D.,1977. Effects of some antimycotics and disinfectants on the Ascosphaera apis Maassen fungus in vitro. Z. angew. Entomology 84: 225-232.
Sanchez O., Navarro R.E., and Aguirre J.,1997. Increased transformation frequency and tagging of developmental genes in Aspergillus nidulans by restriction enzyme-mediated integration (REMI). Molecular Genomics and Genetics 258:89-94.
Sankara K.R., and Prakash A.H.,1995. A simple method for the isolation of plant protoplasts. Journal of Biosciences 20:645-655.
Sato T., Yaegashi K., Ishii S., Hirano T., Kajiwara S., Shishido K., and Enei H.,1998. Transformation of the edible basidiomycete Lentinus edodes by restriction enzyme-mediated integration of plasmid DNA. Bioscience, Biotechnology and Biochemistry 62:2346-2350.
Schiestl R.H., and Petes T.D.,1991. Integration of DNA fragments by illegitimate recombination in Saccharomyces cerevisiae. Proceedings of Natural Academic Sciences, USA 88:7585-7589.
Schillberg S., Tiburzy R., and Fischer R.,2000. Transient transformation of the rust fungus Puccinia graminis f. sp. tritici. Molecular Genomics and Genetics 262:911-915.
Schmid-Hempel P.,1998. Parasites in Social Insects. Princeton University Press, Princeton, New Jersey.
Schmid-Hempel P.,2005. Evolutionary ecology of insect immune defenses. Annual Review of Entomology 50:529-551.
Schulte C., Leboulle G., Otte M., Grtinewald B., Gehne N., and Beye M.,2013. Honey bee promoter sequences for targeted gene expression, Insect Molecular Biology doi: 10.1111/imb.12031
Seong K., Hou Z., Tracy M., Kistler H.C., and Xu J.R.,2005. Random insertional mutagenesis identifies genes associated with virulence in the wheat scab fungus Fusarium graminearum. Phytopathology 95:744-750.
Shah P.A., and Pell J.K.,2003. Etnthomopathogenic fungi as biological control agents. Applied Microbiology and Biotechnology 61:413-423.
Shimanuki H., and Knox D.A.,2000. Diagnosis of Honey Bee Diseases:US Department of Agriculture, Agriculture Handbook No., AH-690:pp 61
Sikandar H., and Christos C.,2010. Isolation and fusion of protoplasts from the phytopathogenic fungus sclerotium rolfsii (sacc). Brazilian Journal of Microbiology 41:253-263
Simlesa V.,2010. Chalkbrood Disease of Honeybees, Agnote No:. Kll, Northern Territory Government, Darwin, pp3.
Skou J.P.,1972. Ascosphaerales. Friesia 10:1-24.
Skou J.P.,1988. More details in support of the class Ascosphaeromycetes. Mycotaxson ⅩⅩⅪ (1): 191-198.
Smith D.R., Villafuerte L., Otisc G., and Palmer M.R.,2000. "Biogeography of Apis cerana F. and A. nigrocincta Smith:insights from mtDNA studies" (PDF). Apidologie 31 (2):265-279.
Spiltoir C.E.,1955. Life cycle of Ascosphaera apis (Pericystis apis), American Journal of Botany 42:501-508.
Spivak M., and Downey D.L.,1998. Field assay for hygienic behavior in honey bees (Hymenoptera:Apidae). Journal of Economic Entomology 91:64-70.
Spivak M., and Gilliam M.,1993. Facultative expression of hygienic behavior in relation to disease resistance. Journal of Apiculture Research 32:147-157.
Stanley D., Yonggyun K., and Youngjin P.,2011. Cellular immunosenescence in adult male crickets, Gryllus assimilis. Archives of Insect Biochemistry and Physiology 76 (4):185-194
Strand M.R.,2008. Insect cellular immunity. Insect Science 15:1-14.
Stroschein-Stevenson S.L., Foley E., O'Farrell P.H., and Johnson A.D.,2009. Phagocytosis of Candida albicans by RNAi-treated Drosophila S2 cells. Methods in Molecular Biology 470: 347-358.
Sugahara M., and Sakamoto F.,2009. "Heat and carbon dioxide generated by honeybees jointly act to kill hornets". Naturwissenschaften 96 (9):1133-1136.
Sunagawa M., Murata H., Miyazaki Y., and Nakamura M.,2007. Transformation of the Mycorrhizal Basidiomycetes, Suillus grevillei and S. bovinus, by Particle Bombardment. Bioscince, Biotechnology and Biochemistry 71(1):47-50.
Suzuki K., Imai Y, Yamashita I., and Fukui S.,1983. In vivo ligation of linear DNA molecule to circular forms in the yeast Saccharomyces cerevisiae. Journal of Bacteriology 155:747-754.
Talkhan F.N., El-Masry H.G., Hassanein S.M., El-Beih F.M. and Abdel-Aziz M.S.,2008. Genetic improvement of dextranase Production by Penicillium funiculosum via Mutation Induction and Protoplast Fusion. International Journal of Agriculture and Biology 10(1):42-46.
Tamai Y., Miura K., and Kayama T.,1990. Electrical fusion of protoplasts between two Pleurotus species. Mokuzai Gakkaishi 36:487-490.
Tanaka A., Shiotani H., Yamamoto M., and Tsuge T.,1999. Insertional mutagenesis and cloning of the genes required for biosynthesis of the host-specific AK-toxin in the Japanese pear pathotype of Alternaria alternata. Molecular Plant Microbe Interaction 12:691-702.
Teshome G.,2006. Characterization of the causative agent of Chalkbrood disease of honeybees (Apis mellifera L.) in Ethiopia, MSc. Thesis, Addis Ababa University, School of Graduate Studies, Department of Biology, Addis Ababa, Ethiopia, pp61.
Thawley C.H.,2010. "Heat tolerance as a weapon". Davidson College. Retrieved June 1,2010.
Theantana T., and Chantawannakul P.,2008. Protease and β-N-acetylglucosaminidase of honey bee chalkbrood pathogen Ascosphaera apis, Journal of Apicultural Research 1:68-76.
Thon M.R., Nuckles E.M., and Vaillancourt L.J.,2000. Restriction Enzyme-Mediated Integration Used to Produce Pathogenicity Mutants of Colletotrichum graminicola. Molecular Plant Microbe Interation 13(12):1356-1365.
Tilburn J., Scazzocchio C., Taylor GG, Zabicky-Zissman J.H., Lockington R.A., and Davies R.W., 1983. Transformation by integration in Aspergillus nidulans. Genetics 26(2-3):205-221.
Timberlake W.E., and Marshall M.A.,1989. Genetic engineering of filamentous fungi. Science. 244:1313-1317.
Toyomatu T.M., and Mori N.D.K.,1986. Interspecific protoplast fusion between Pleurotus ostreatus and Pleurotus salmoneo-stramineus. Agricultural Biology and Chemistry 50: 223-225.
Tunlid A., Ahman J., and Oliver R.P.,1999. Transformation of the nematode-trapping fungus Arthrobotrys oligospora. FEMS Microbiological Letters 173:111-116.
Tutkun E., Maden S., Inci A., and Yilmarz B.,1993. General situation of chalkbrood disease in honeybees in Turkey. Turk Entomol Derg-TU 17 (2):65-68.
Vandenberg J.D.,1992. Bioassay of the Chalkbrood Fungus Ascosphaera aggregata on Larvae of the Alfalfa Leafcutting Bee, Megachile rotundata. Journal of Invertebrate Pathology 60: 159-163.
vanEngelsdorp D., and Marina D.M.,2010. A historical review of managed honey bee populations in Europe and the United States and the factors that may affect them. Journal of Invertebrate Pathology 103:S80-S95
Vojvodic S., Jensen A.B., Markussen B., Eilenberg J., and Boomsma J.J.,2011. Genetic Variation in Virulence among Chalkbrood Strains Infecting Honeybees, PLoS ONE 6(9):e25035.
Vojvodic S., Jensen A.B., James R.R., Boomsma J.J., and Eilenberg J.,2011. Opposing temperature-dependent mortality after infection with an obligate and facultative fungal pathogen of honeybees. Veterinary Microbiology 149:200-205.
Wang Y., Guo B., Miao Z.Q., and Tang K.,2007. Transformation of taxol-producing endophytic fungi by restriction enzyme-mediated integration (REMI), FEMS Microbiological Letters 273: 253-259.
Warwar V., Oved S., and Dickman M.B.,2000. Antisense expression of the calmodulin gene from Colletotrichum trifolii impairs prepenetration development. FEMS Microbiological Letters 191:213-219.
Wilson W.T., Nunamaker R.A., and Maki D.,1984. The occurrence of brood diseases and the absence of the Varroa mite in honeybees in Mexico. American Bee Journal 124:51-53.
Wood M.,1998. Microbes help bees battle chalkbrood. Journal of Economic Entomology 16-17.
Wubie A.J., Hu Y-Y., Li W., Huang J-X., Guo Z., Xu S-F., and Zhou T.,2013. Factors analysis in protoplast isolation and regeneration from a chalkbrood fungus, Ascosphaera apis. International Journal of Agriculture and Biology 16:89-96.
Yacobson B.A., Elad D., Rosenthal K., Kramer I., Slovecky I., and Efrat H.,1991. A recent chalkbrood outbreak in Israel:attempts at therapeutic intervention. American Bee Journal 131: 786.
Yakoby N.s Zhou R., Kobiler I., Dinoor A., and Prusky D.,2000. Development of Colletotrichum gloeosporioides Restriction Enzyme-Mediated Integration Mutants as Biocontrol Agents Against Anthracnose Disease in Avocado Fruits. The American Phytopathological Society Vol. 91, No.2,143-148. Publication no. P-2000-1218-01R
Zaghloul O.A., Mourad A.K., El Kady M.B., Nemat F.M., and Morsy M.E.,2005. Assessment of losses in honey yield due to the chalkbrood disease, with reference to the determination of its economic injury levels in Egypt. Communications in Agricultural and Applied Biological Sciences 70 (4):703-714.
Zhao K., Zhou D., Ping W., and Jingping G.,2004. Study on the Preparation and Regeneration of Protoplast from Taxol-producing Fungus Nodulisporium sylviforme; Nature and Science 2(2): 52-59.
Zhou X., Yamin W., Huifang Z., Zinan W., Juan L., Lu L., and Kexuan T.,2008, Protoplast formation, regeneration and transformation from the taxol-producing fungus Ozonium sp., African Journal of Biotechnology 7 (12):2017-2024.
Zhou X.W., Wang Z.N., Wei Y.M., Jiang K.J., Lin J., Sun X.F., Tan F., Miao Z.Q., and Tang K.X., 2006. Selection of optimal fermentation medium and enzyme system for isolation of protoplasts from endophytic fungus, Taxus mairei. Chin. Journal of Applied Environment and Biology 12(2):176-181.
Zhou X.Y., Xu S., Liu L., and Chen J.,2007. Degradation of cyanide by Trichoderma mutants constructed by restriction enzyme mediated integration (REMI). Bioresource Technology 98: 2958-2962.