分子昆虫学实验室

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 一、研究方向:

 (一)  昆虫中肠基因和蛋白表达谱的研究

对斜纹夜蛾中肠不同发育时期的基因表达进行了基因表达顺序标签的分析,共得到6815ESTs并拼接组装成3037个单一基因。鉴定出大量与糖代谢、蛋白代谢、脂肪代谢和核酸代谢与免疫、抗病性相关的基因。同时,我们对中肠蛋白表达也进行了分析,利用2 维电泳和质谱方法分析了斜纹夜蛾在5 个发育时期中肠蛋白的差异表达。发现至少60 多个低丰度的蛋白在不同时期发生可检测的明显变化。利用Shotgun质谱蛋白分析方法, 对进食期中肠蛋白进行了分析, 发现2043种蛋白肽段, 鉴定出与糖、脂肪酸、蛋白和氨基酸与核酸运输和代谢相关的蛋白。全面系统的基因和蛋白表达分析为我们了解中肠在不同发育时期的功能变化奠定坚实的基础,并为鉴定作用于中肠的分子靶标提供可能。

(二)  功能基因的克隆与分析

昆虫是不能自主地合成甾醇的,必须从食物中吸收。甾醇在中肠的吸收是通过甾醇运输蛋白实现的。我们从斜纹夜蛾中克隆了这个基因并对其在不同组织和发育时期的表达,组织细胞定位和功能进行了分析。结果表明,斜纹夜蛾甾醇运输蛋白SCPx是中肠专一表达的蛋白,它可能通过吸收和运输固醇类物质影响昆虫的生长发育。这是第一个经详细研究证明其生理功能的鳞翅目昆虫甾醇运输蛋白。目前我们已得到该重组蛋白的结晶, 正对其结构和功能及其调节进行深入的研究。

昆虫在进食时,中肠要消化分解食物中的蛋白和脂肪成小分子的氨基酸和脂肪酸后,后者才被吸收进入中肠细胞内供中肠或其它组织细胞生长需要。 我们对斜纹夜蛾中肠的丝氨酸蛋白酶和脂肪酸结合蛋白进行了大量的研究。已克隆得到2个不同的丝氨酸蛋白酶和8个脂肪酸结合蛋白基因的全长cDNA,并对它们的表达、定位和功能进行了分析。结果表明,这些蛋白均与昆虫进食有关。这些工作为我们今后利用丝氨酸蛋白酶和脂肪酸结合蛋白为靶标抑制昆虫生长打下的基础。

我们还对中肠中两个新的未知功能的基因(Un17Un7)进行了较为深入的分析,以确定其在昆虫中肠中的生理学作用。这是两个尚未见任何报道的昆虫新基因。

(三)  昆虫变态发育的研究

我们对家蚕蛹期专一表达的一个翅膀几丁质蛋白的表达的分子调控机理进行了深入的研究。这个基因是蛹期专一表达的。我们发现, 这个基因受到蜕皮激素正调控, 而保幼激素可以抑制蜕皮激素的作用。我们鉴定出该基因的两个调节元件,分别受转录因子 BmPOUM2BmbFTZ-F1的调控。 蜕皮激素正是通过这两个调控因子调节了翅膀几丁质蛋白的表达,使到幼虫能向蛹期发育。我们还对不同发育时期与变态相关的系列基因的表达进行了分析。

(四)  昆虫抗药性研究

昆虫抗药性调节机理的研究对农业害虫防治有着重要的理论和实践意义。昆虫对杀虫剂产生抗性的一个重要机制是谷胱甘肽-S-转移酶(GSTs)解毒作用增强。我们已从斜纹夜蛾获得了8个斜纹夜蛾GSTs cDNA,并已对其中几个基因的结构、启动子的结构和调控、基因表达和酶活性和动力学以及抗药性等进行了初步分析。正进一步对这些GSTs基因表达调节与昆虫抗药性机理作更深入的分析。

 二、研究课题:

²  国家级课题项目:

1.  分子靶标导向的绿色化学农药创新研究,科技部973计划项目, 2009-2013,冯启理参加子课题 。

2.  家蚕主要经济性状功能基因组与分子改良研究(2005CB121002),科技部973计划项目,冯启理参加子课题:昆虫激素对蛹期专一表达基因的调控机理研究

3.  家蚕重要经济性状功能基因的克隆、分子调控与应用研究(2006AA10A119),国家高技术研究发展计划(863计划)项目, 2006-2010,冯启理参加子课题。

4.  斜纹夜蛾中肠分子生物学和功能基因组学研究30671408),  国家自然科学基金面上项目, 2007-2009,冯启理主持。

5.  丝氨酸蛋白酶及其抑制因子在昆虫蜕皮和变态发育中的作用30571258),国家自然科学基金面上项目,2006-2008,冯启理主持。

6.  昆虫生长发育过程中热激蛋白基因与蜕皮激素的相互关系研究(30900152),国家自然科学基金青年科学基金项目, 2009-2012,黄立华主持。

²  省级课题项目:

1.       斜纹夜蛾中肠基因EST分析和功能基因克隆研究(06105204),广东省自然科学基金重点项目,2006-2009 冯启理主持。

2.       昆虫蜕皮和变态发育的分子生物学研究(5005941),广东省自然科学基金面上项目,2006-2007,冯启理主持。

3.       斜纹夜蛾谷胱甘肽-S-转移酶基因抗性机理的研究(9151063101000009),广东省自然科学基金面上项目,2009-2012,郑思春主持

4.       斜纹夜蛾变态相关基因的鉴定及其功能分析 9451063101002022),广东省自然科学基金项目博士启动基金, 2009-2012黄立华主持

5.       卵母细胞特异生长因子GDF9在斑马鱼性腺分化和发育中的作用 845106310000345),  广东省自然科学基金项目博士启动基金, 2008-2010,刘琳主持。

 

²  横向课题项目及其它:

1.       杆状病毒和宿主细胞间的相互作用(2008AA000238),科技部国际科技合作与交流专项-中荷科学战略联盟项目,2008-2013,冯启理参加子课题。

2.       家蚕重要经济性状功能基因的克隆、分子调控与应用研究--家蚕蛹期发育变态的分子调控研究,国家人事部留学人员科技活动项目择优资助经费, 2007-2010,冯启理主持。

3.       国际昆虫中肠生物学研讨会,国家自然科学基金委对外交流与合作项目(30671408), 2008,冯启理主持。

4.       国际昆虫中肠生物学研讨会,广东省科技厅对外交流与合作处项目 306714082008,冯启理主持。

5.       热激蛋白基因在斜纹夜蛾抗寒性、生长发育和生殖中的作用机理研究(ChineseIPM0902),农业虫害鼠害综合治理研究国家重点实验室开放课题, 2009-2010,黄立华主持。

 

 三、发表文章(2005~)

  1. Liu, J.S. Zheng, S.C., Liu, L., Li, L., Feng, Q.L.* 2010. Protein profiles of the midgut of Spodoptera litura larvae at the sixth instarfeeding stage by shotgun ESI-MS approach. Journal of Proteome Research 9: 2117-2147.(IF: 5.684)

  2. Ming-Fang He, Lin Liu , Wei Ge, Pang-Chui Shaw, Renwang Jiang , Li-Wha Wu, and Paul Pui-Hay But.  2009. Antiangiogenic activity of Tripterygium wilfordii and its terpenoids. Journal of Ethnopharmacology   Jan 12;121(1):61-8

  3. Yufen Huang, Peter J. Krell, Tim Ladd, Qili Feng, Sichun Zheng*. 2009. Cloning and expression of two glutathione S-transferase cDNAs in the spruce budworm, Choristoneura fumiferana. Arch Insect Biochem Physiol. 70(1):44-56

  4. Huimin Deng, Yufen Huang, Qili Feng, Sichun Zheng*  2009. Two epsilon glutathione S-transferase cDNAs from the common cutworm, Spodoptera litura: Characterization and developmental and induced expression by insecticides J Insect Physiol. 55 (12):1174-1183

  5. Xing-Rong Guo, Si-Chun Zheng, Lin Liu, Qi-Li Feng* 2009. The sterol carrier protein 2/3-oxoacyl-CoA thiolase (SCPx) is involved in cholesterol uptake in the midgut of Spodoptera litura: gene cloning, expression, localization and functional analyses. BMC Mol. Biol 10(1):102-120.

  6. Wen-Ying He, Yi-Ping Zheng, Lin Tang, Si-Chun Zheng, Catherine Beliveau, Daniel Doucet, Mchel Cusson and Qi-Li Feng* 2009. Cloning, expression and localization of a trypsin-like serine protease in the spruce budworm, Choristoneura fumiferana Insect Science 16:455-464

  7. Shui-Kei Poon, Wai-Kin So, Xiaobin Yu, Lin Liu and Wei Ge*  (2009) Characterization of Inhibin α Subunit (inha) in the Zebrafish – Evidence for Differential Functions of Pituitary FSH and LH in the Ovary and a Potential Feedback Loop between the Pituitary and Ovary. Reproduction 138 (4): 709-719

  8. Y.-P. Zheng, W.-Y. He, C. B´eliveau, A. Nisole, D. Stewart, S.-C. Zheng, D. Doucet, M. Cusson, Q.-L. Feng* 2009. Cloning, expression and characterization of four serpin-1 cDNA variants from the spruce budworm, Choristoneura fumiferanag Comp. Biochem Physiol B Biochem.Mol Biol 154(2):165-73

  9. Doucet D, Béliveau C, Dowling A, Simard J, Feng Q, Krell PJ, Cusson M. 2008.  Prophenoloxidases 1 and 2 from the spruce budworm, Choristoneura fumiferana: molecular cloning and assessment of transcriptional regulation by a polydnavirus. Arch Insect Biochem Physiol. 67(4):188-201.

  10. Zheng YP, Krell PJ, Doucet D, Arif BM, Feng Q.L.* 2008. Cloning, expression, and localization of a molt-related beta-N-acetylglucosaminidase in the spruce budworm, Choristoneura fumiferana. Arch Insect Biochem Physiol. 68(1):49-59.

  11. Sichun Zheng,Huimin Deng, Tim Ladd, Bill L.Tomkins,  Peter  J. Krell, Qili Feng*. 2007, Clonging and Characterization of Two Glutathione S-Transferase cDNAs in the Spruce Budworm, Choristoneura fumiferana. Arch. Insect Biochem Physiol. 66(3):146-157.   

四、会议论文  

  1. Qili Feng Functional genomics of lipid and fatty acid metabolisms in the midgut of Spodoptera litura The second International Symposium on Insect Physiology, Biochemistry and Molecular Biology 2009:31 ChengDe, Hebei, China, July 24-28,2009 (invited presentation)

  2. Qili Feng Molecular regulation of the expression of a cuticle protein gene in the silkworm, Bombyx mori International symposium on Bombyx mori functional genomics and modern silk road. Chongqin, Oct. 2009 (Invited presentation)

  3. Qili Feng Can we keep a pupa staying in a cocoon for a longer time? The 6th Asia-Pacific Congress of Entomology (APCE) Beijing Oct. 2009 (Invited presentation)

  4. Qili Feng Identification and characterization of sterol carrier protein (SCPx) as a midgut-specific target for novel insecticides in Spodoptera litura The first China-Japan-Korea workshop on pesticide science Shanghai 70 Nov.2009 (Oral presentation)

  5. Qili Feng Cloning, characterization, expression, localization and functional analysis of a chymotrypsin-like serine protease cDNA in the common cutworm, Spodoptera litura The first China-Japan-Korea workshop on pesticide science Shanghai 80 Nov.2009 (poster)

  6. Qili Feng Hormonal regulation of the expression of a pupa-specific cuticle protein gene through its POU cis-regulatory element 2009年美国昆虫学大会 美国Indianapolis Dec.,2009 (Oral presentation)

  7. Huimin Deng, Qili Feng, Sichun Zheng* Molecular Characterization and Regulation of Cuticle Protein Gene BmWCP4 in the Silkworm, Bombyx mori The Second International Symposium on Insect Physiology, Biochemistry and Molecular Biology 2009:30 (oral presentation)

  8. Yufen Huang, Qili Feng Sichun Zheng* Cloning and characterization of glutathione S-transferase genes from the common budworm, Spodoptera litura The Second International Symposium on Insect Physiology, Biochemistry and Molecular Biology 2009:50 (poster)

  9. Zhiqiang Huang, Daohua Zhou Sichun Zheng, Qili Feng, Lin Liu* Cloning, expression and ligand-binding activity of a midgut-specific fatty acid-binding protein in Spodoptera litura The Second International Symposium on Insect Physiology, Biochemistry and Molecular Biology 2009:50 (poster)

  10. Mingjiong Liao Qili Feng* Preparation of chlorpyrifos microcapsules in gelatin and sodium alginate The Second International Symposium on Insect Physiology, Biochemistry and Molecular Biology 2009:51 (poster)

  11. Xinhua Yang, Sichun Zheng Qili Feng* Responses of ecdyteroid receptors and transcription factors to 20E during metamorphosis in the epidermis, midgut and fat body of the silkworm, Bombyx mori. The Second International Symposium on Insect Physiology, Biochemistry and Molecular Biology 2009:51 (poster)

  12. Qinglei Zhan, Sichun Zheng, Qili Feng, Lin Liu* Cloning, characterization, expression and functional analysis of a midgut-specific chymotrypsin Gene (SlTC1) from Spodoptera litura The Second International Symposium on Insect Physiology, Biochemistry and Molecular Biology 2009:52 (poster)

  13. Chun Zhang, Sichun Zheng, Qili Feng* A midgut-specific chymotrypsin-like serine protease cDNA is associated with food digestion in the common cutworm Spodoptera litura The Second International Symposium on Insect Physiology, Biochemistry and Molecular Biology 2009:52 (poster)

  14. Chun Zhang, Sichun Zheng, Qili Feng* A chymotrypsin-like serine protease cDNA involved in food protein digestion in the common cutworm, Spodoptera litura: The 6th Asia-Pacific Congress of Entomology (APCE) Beijing Oct. 2009 (Oral presentation)

  15. Qili Feng, Gene and protein expression profiles in the midgut of Spodoptera litura during molting and metamorphosis. International Symposium on Insect Midgut Biology, Guangzhou, China, April 7-11, 2008.

  16.  Wenyin He, Identification of midgut-specific genes expressed in the Spodoptera litura midgut. International Symposium on Insect Midgut Biology, Guangzhou, China, April 7-11, 2008.

  17.  Daohua ZhouAnalysis of expressed sequence tags from the midguts of Spodoptera litura (Lepidoptera: Noctuidae) at 5th to 6th molting, feeding 6th instar and prepupal stages. International Symposium on Insect Midgut Biology, Guangzhou, China, April 7-11, 2008.

  18. Jisheng LiuProtein expression profiles in the midgut of Spodoptera litura during molting and metamorphosis or feeding on different host plants. International Symposium on Insect Midgut Biology, Guangzhou, China, April 7-11, 2008.

  19.  Xinrong Guo, Cloning and identification of sterol carrier protein cDNA from the midgut of Spodoptera litura. International Symposium on Insect Midgut Biology, Guangzhou, China, April 7-11, 2008.

  20.  Mingjiong Liao, Ultrastructural changes and programmed cell death of the midgut epithelium of Spodoptera litura during molting and metamorphosis. International Symposium on Insect Midgut Biology, Guangzhou, China, April 7-11, 2008.

五、教学

 本室开设课程:

    《昆虫生理与分子生物学》、《分子生物学实验技术》、《昆虫基因组学》、《细胞生物学研究进展》