冰核细菌对枇杷幼果受冻致萎的影响
|
摘要
霜冻在我国发生区域广，危害的植物种类多，是一种严重的农业气象灾害。本文以南方主栽树种枇杷为研究对象，采用定期定量分离的方法，检测INA细菌在枇杷果实形成过程中的数量分布，并研究同一生长环境下不同枇杷品种之间INA细菌消长动态变化，以及不同果园环境下(对应温度、降雨量、结露、紫外线照射强度、海拔高度和微生物种类等不同因素)，对INA细菌消长动态变化的影响及其互动互作关系，以期查明INA细菌在枇杷上的消长动态规律。主要结果如下：
1、查明了福建枇杷树上INA细菌种类
从福建地区枇杷树上分离到24株INA细菌，经细菌学鉴定，初步确定为凤梨欧文氏菌。
2、初步明确了枇杷树上INA细菌消长动态规律
大量调查研究结果表明，INA细菌分布广泛。在枇杷花期就已经有INA细菌的存在，并且INA细菌在枇杷不同部位上的分布存在差异，差异明显：不同品种间也存在差异，差异不明显。
采用定性定量和定期取样分离方法，对福建莆田地区早钟6号进行了INA细菌消长动态规律研究，结果表明，INA细菌密度在田间枇杷树上有2个高峰期：花序分化期(9月中旬至11月初)，果实膨大期(12月初至3月中旬)：其他季节(炎热的夏季)数量少，甚至难以分离到。
INA细菌分布数量与枇杷品种、地理位置及生态条件有关，在幼果上易分离到INA细菌，其最高密度为1.13×10~7cfu／g鲜重，表明在幼果期INA细菌分布数量多，易遭受霜冻危害，故为除INA细菌防御霜冻提供了科学依据。
3、验证了INA细菌能够诱发和加重枇杷幼果冻害
人工气候室及田间试验结果都表明，接种处理的枇杷幼果冻害严重度比同一温度下未接种INA细菌的高，加重了枇杷幼果霜冻，再次证明INA细菌是诱发和加重枇杷幼果霜冻害的重要因素。
In this paper, the species, distribution and dynamic of INA on loquat were reported as well as the screening of bactericides for forest control in Fujian District. These results were illustrated as follows:
First, the species of INA bacteria on loquat in Fujian were identified.Twenty-four strains of INA bacteria were isolated from loquat. Based on bacteriological methods, they were identified belonging to one genera: E.ananas.
Second, their distribution and dynamic of INA bacteria was demonstrated primarily.
By investigation, INA bacteria could be isolated from most of loquat, showing INA bacteria were widespread in loquat. The distribution of INA bacteria varied in loquat cultivar, geographic position and ecological conditions. They were easily isolated and had a maximum populations 1.13×10~7CFU/g.f.w.
In Putian zone, INA bacteria population on "Zaozhong", was checked periodically. The results revealed two peaks of population during flowering stage and fruiting stage. On the contrary. INA bacteria could not be isolated easily during hot summer. It was the first reported that populations of INA bacteria varied in seasons.
Third, it was testified that INA bacteria could induce and aggravate frost injury on loquat.
The frost index of loquat after application of INA bacteria was higher than that of loquat treated with distilled water in programmable frost chamber; Similar examples happened in field. All the results illustrated that INA bacteria could induce and aggravate frost injury on loquat.
1、查明了福建枇杷树上INA细菌种类
从福建地区枇杷树上分离到24株INA细菌，经细菌学鉴定，初步确定为凤梨欧文氏菌。
2、初步明确了枇杷树上INA细菌消长动态规律
大量调查研究结果表明，INA细菌分布广泛。在枇杷花期就已经有INA细菌的存在，并且INA细菌在枇杷不同部位上的分布存在差异，差异明显：不同品种间也存在差异，差异不明显。
采用定性定量和定期取样分离方法，对福建莆田地区早钟6号进行了INA细菌消长动态规律研究，结果表明，INA细菌密度在田间枇杷树上有2个高峰期：花序分化期(9月中旬至11月初)，果实膨大期(12月初至3月中旬)：其他季节(炎热的夏季)数量少，甚至难以分离到。
INA细菌分布数量与枇杷品种、地理位置及生态条件有关，在幼果上易分离到INA细菌，其最高密度为1.13×10~7cfu／g鲜重，表明在幼果期INA细菌分布数量多，易遭受霜冻危害，故为除INA细菌防御霜冻提供了科学依据。
3、验证了INA细菌能够诱发和加重枇杷幼果冻害
人工气候室及田间试验结果都表明，接种处理的枇杷幼果冻害严重度比同一温度下未接种INA细菌的高，加重了枇杷幼果霜冻，再次证明INA细菌是诱发和加重枇杷幼果霜冻害的重要因素。
In this paper, the species, distribution and dynamic of INA on loquat were reported as well as the screening of bactericides for forest control in Fujian District. These results were illustrated as follows:
First, the species of INA bacteria on loquat in Fujian were identified.Twenty-four strains of INA bacteria were isolated from loquat. Based on bacteriological methods, they were identified belonging to one genera: E.ananas.
Second, their distribution and dynamic of INA bacteria was demonstrated primarily.
By investigation, INA bacteria could be isolated from most of loquat, showing INA bacteria were widespread in loquat. The distribution of INA bacteria varied in loquat cultivar, geographic position and ecological conditions. They were easily isolated and had a maximum populations 1.13×10~7CFU/g.f.w.
In Putian zone, INA bacteria population on "Zaozhong", was checked periodically. The results revealed two peaks of population during flowering stage and fruiting stage. On the contrary. INA bacteria could not be isolated easily during hot summer. It was the first reported that populations of INA bacteria varied in seasons.
Third, it was testified that INA bacteria could induce and aggravate frost injury on loquat.
The frost index of loquat after application of INA bacteria was higher than that of loquat treated with distilled water in programmable frost chamber; Similar examples happened in field. All the results illustrated that INA bacteria could induce and aggravate frost injury on loquat.
引文
[1] 何维勋．霜冻[M]．北京：农业出版社，1986．272～274．
[2] 罗心平，尼章光，王跃全，等．蛋黄果引种试种研究初报[J]．热带农业科技，2004，27(1)：47～48．
[3] 尼章光，罗心平，王跃全，等．值得开发的热带亚热带果树[J]．中国南方果树，2003，32(1)：25～26．
[4] 肖鸿修．中国热带南亚热带果树[M]．北京：中国农业出版社，1998．341～342．
[5] 冯玉香，何维勋．细菌冰核提高印度谷螟过冷却点的研究[J]．昆虫学报，1996，39(1)：53～57．
[6] 孙忠富．霜冻灾害与防御技术[M]．北京：中国农业出版社，2001．10～15．
[7] 唐广，蔡涤华，郑大玮．果树蔬菜霜冻与冻害的防御技术[M]．农业出版社，1993．30～57．
[8] 冯玉香，何维勋，崔松林．霜冻时冰晶在作物体内生长的试验研究[J]．中国农业气象，1991，12(4)：6～9．
[9] LEVITT, J.Freezing temperature responses of plants to environmental stress[M]. New York: Academic Press. 1979, 1: 267～291.
[10] PALTA, J.P., JEMSEM, et al. Cell membrane alterations following slow free thaw, injury and recovery[M]. Plant Cold Hardiness and Freezing Stress. New York: Academic Press, 1982, 2: 221～242.
[11] 何维勋，冯玉香，夏满强．解冻速率对农作物霜冻害的影响[J]．应用气象学报，1993，4(4)：440～445．
[12] 郭修武，傅望衡．葡萄根系抗寒性的研究[J]．园艺学报，1989，16(1)：17～22．
[13] 杨建民，李艳华，杨敏生，等．几个仁用杏品种抗寒性的比较[J]．中国农业科学，1999，32 (1)：46～50．
[14] 王飞，陈文登，高爱琴，等．杏品种一年生休眠枝、花、幼果抗寒的相关分析[J]．西北植物学报，1999，(4)：618～622．
[15] 彭伟秀，杨建民．杏花器官组织抗寒性研究[J]．果树学报，2002，19(2)：108～110．
[16] 白琳，白靳来．杏树花期、幼果期抗寒力调查[J]．北方园艺，1992，2(6)：69～70．
[17] 孙福在，朱红，何礼远．影响冰核细菌成冰活性的因素研究[J]．中国农业科学．1991，24(3)：57～64．
[18] 孙福在．我国生物冰核研究进展[J]．中国农业科学．1996，29(5)：62～68．
[19] OBATA, H., TAKINAMI, K, TANISHITA, J.T., et al. Identification of a new nucleation bacterium and its ice nucleation properties[J]. Agric Biol Chem, 1990, 54: 725～730.
[20] ORSER, C., STASKAWICZ, B.J., LOPER, J. et al. Cloning of genes involved in bacterial ice nucleation and fluorescent pigment production[J]. Agric Biol Chem, 1983, 12: 353～361.
[21] 高桥辛吉．INA细菌研究现状用化[J]．冷冻(日)，1987，62(718)：73～80．
[22] POOIEY, L., BROWN, T.A. Preparation of cell-free ice nuclear from Pseudomonas syringe[J]. Proc R Soc Lond Set B Biol Sci, 1990, 241: 112～115.
[23] 刘建华，陶毓汾．冰核细菌与玉米和大豆霜冻关系的研究[J]．中国农业气象，1990，11(2)：1～6．
[24] EDWARD, A.R., VANDENBUSSCHE, R.A., WICHMAN, H.A., et al. Unusual pattern of bacterial ice nucleation gene evolution[J]. Mol Biol Ecol, 1994, 11: 911～920.
[25] 杨建民，周怀军．果树霜冻害研究进展[J]．河北农业大学学报，2000，23(3)：54～58．
[26] 孙福在，赵廷昌．杏树上冰核细菌种类及冰核活性与杏花霜冻关系的研究[J]．中国农业科学，2000，(6)：50～58．
[27] LINDOW S E. The role of bacteria ice nucleation in frost injury to plant[J]. Ann. Rev. Phytopathol. 1983, 21: 326～384.
[28] ARAI, S. WATANABE, M.Freeze texturing of food materials by ice-nucleation with the bacterium Erwinia ananas[J]. Agric. boil. Chem., 1986, 50(1): 169～175.
[29] 杨建民，孟庆瑞．冰核细菌对杏花抗寒性的影响[J]．园艺学报，2002，29(1)：20～24．
[30] 韩伟秀，杨建民．冰核细菌对仁用杏花粉超微结构的影响[J]．园艺学报，2001，(5)：453～456．
[31] 孙福在，赵廷昌．冰核细菌生物学特性及其诱导植物霜冻机理与防霜应用[J]．生态学报．2003，23(2)：336～346．
[32] 张世光．PCR在快速检测冰核细菌及其冰核活性强度中的应用[A]．海峡两岸植物病理学术研讨会会刊[C]．中华植物病理学会，1997，4．
[33] PHELPS P, GIDDINGS TH, PHOCHODA M, et al. Release of cell-free ice nuclei by Erwinia herbicola[J]. Bactedol, 1986, 167: 496～502.
[34] MASANORI, KOIKE. Random Amplified Polymorpruz DNA Analysis of Japanese Isolates of verticilitun dahliae and Valbo-atrum[J]. Plant Disease, 1996, 80: 1224～1227.
[35] BASSI, A.S., DING, D.N., GLOOR, G.B., et al. Expression of a single chain antibodies(ScFVs) for c-myc oncoprotein in recombinant Escherichia coli membranes by using the ice-nucleation protein of Pseudomonas syringae[J]. Biotechnol. Prog., 2000, 16: 557～563.
[36] 唐朝荣．冰核细菌Erwinia ananas 110冰核基因克隆、序列分析和促冻杀虫基因工程菌构建[D]．中国农业科学院博士论文，2001．
[37] 唐朝荣，孙福在，赵廷昌，等．冰核细菌Erwinia ananas 110冰核基因克隆和序列分析[J]．微生物学通报，2002，29(4)：45～51．
[38] BIGG, E.K. The supercooling of water[J]. Proc Phys Soc B, 1953, 66: 688～694.
[39] ORSER C., STASKAWICZ, B.J., LOPER, J. et al. Cloning and expression of bacterial ice nucleation genes in Escherichia coli[J]. Bacterial. 1985, 164: 359-366.
[40] 姬广海，张世光．冰核活性细菌冰核基因多态性研究[J]．云南农业大学学报，1997，12(3)：153～157．
[41] WARREN, GJ., COROTTO, L., WOLBER, P.Conserved repeats in diverged ice nucleation structural genes from two species of Pseudomonas[J]. Nucl Acids Res, 1986, 20: 8047～8060.
[42] WOLBER, P., WARREN, G.J. Bacterial ice nucleation proteins[J]. TIBS, 1989, 14: 179.
[43] 任修海．冰核细菌的生物学及生物技术开发[J]．生物技术通报，1993，(6)：12～15．
[44] KOZLOFF, L.M., SCHOFIELD, et al. Ice nucleating activity of Psedomonas syringae and Erwinia herbicola[J]. Baceteriol., 1983, 153: 222～231.
[45] SCHNELL, R.C., VALI,G. Wodd-wide source of leaf-derived freezing nucIei[J]. Nature, 1973, 246: 212～213.
[46] WATANABE. M, ARAI.S. Bacterial ice nucleation activity and its application to freeze concentration of fresh food for modification of their properties[J]. Food Eng, 1994, 22: 453～473.
[47] LEEUNG, KWON MIKYUNG, SEONG KIYONG, et al. Development of a specific antibody for the detection of ice nucleation-active bacteria[J]. Plant Disease and Agriculture, 1999, 5(1): 27～33.
[48] TURNER, ARELLANOF, KOZLOFFM. Components of ice nucleation structures of bacteria[J]. Bacterial, 1991, 173: 6515～6527.
[49] 孙福在，朱红，何礼远，等．冰核细菌对光肩星天牛幼虫促冻杀虫研究[J]．林业科学研究，1997，10(1)：96～99．
[50] 张耀东，曹阳．冰核活性细菌对储粮害虫耐冻性的影响[J]．郑州粮食学院学报，1998，19(2)：1～5．
[51] 杨建民．INA细菌与杏树花器官霜冻害关系研究[D]．中国农业大学博士论文，2002，59-73．
[52] 王凤才．对2001年“三·二八”果树特大冻害的评估分析[J]．落叶果树，2002，3：11～14．
[53] 吕增仁．我国杏研究进展[J]．河北果树，1996，29(5)：62～67．
[54] 冯玉香．黄瓜霜冻与冰核细菌活性细菌的关系[J]．园艺学报，1990，17(3)：211～216．
[55] 何维勋，冯玉香，孙福在，等．防御霜灾新途径的研究[J]．灾害学，1990，(1)：14～19．
[56] 冯玉香，何维勋，夏满强．作物霜冻与低温强度及冰核菌密度的关系[J]．应用气象学报，1995，6(1)：90～91．
[57] 韦建福．我国生物冰核研究进展[A]．植物病虫害生物学研究进展一植物病虫害生物学国家重点实验室研究论文选[C]．北京：中国农业科技出版社，1995．
[58] ARNY D.C., LINDOW S E, UUPPER C D. Frost sensitivity of Zea may be increased by application of Pseudomnas Syingae[J]. Nature, 1976, 266: 282～284.
[59] LINDOW S E. Competitive exclusion of epiphytic bacteria by ice-mutants of Pseudomonas syringae[J]. Appl and Environ Microbiol, 1987, 53: 2520～2527.
[60] 朱红，孙福在，张永祥．冰核细菌对棉铃虫结冰温度影响的研究[J]．中国农业科学，1994，27(6)：23～27．
[61] 方中达．植病研究方法(第三版)[M]．北京：中国农业出版社，1998．
[62] 孙福在，赵廷昌，何维勋，等．应用生防菌和药剂防除冰核细菌控制玉米霜冻研究[M]．北京：中国农业科技出版社，2001．
[63] 张世光．云南冰核细菌的鉴定及防霜新技术研究[J]．云南农业大学学报，1995，10(2)：192～197．
[64] KIM, K.C., LEE, U., SONG, D.U. et al. Purification and characterization of ice nucleation-active bacteria[J]. Korean L Plant Pathol, 1996, 12(1): 99～108.
[65] WOLBE, P.K. Bacterial ice nucleation[J]. Adv. Microb. Physiol., 1993, 34: 203～207.
[66] WISNIEWSKIM., GLENN, D.M., FULLER, M.P.Use of a hydrophobic particle film as a barrier to extrinsic ice nucleation in tomato plants[J]. Hort Sci, 2002, 127(3): 358～364.
[67] 杨丽．冰核细菌活性的冰核基因克隆及其在大肠杆菌中表达[J]．中国农业科 学，1993，26(5)：88～89．
[68] LINDOW, S.E. Control of epiphytic ice nucleation-active bacteria for management of plant frost injury[M]. In: Biological Ice Nucleation and its Applications. Lee RE, Warren G and Gusta LV(eds). APS Press. 1996.
[69] GARETHE, WARREM. Bacteria Ice Nucleation: Molecular Biology and Application[J]. Biotechnology and Genetic Engineering Review, 1987, 5 (7): 88～89.
[70] LI, J.K., LEE, J.C. Bacterial ice nucleation and its potential application on the food industry. Trend Food Sci, 1995, 6: 259～263.
[71] 任修海．成冰核细菌的生物学及生物技术开发[J]．生物技术通报，1993，(6)：12～15．
[72] WARD P.J., DEMOTT P.J. Preliminary experimental valuation of SNOMAX snow inducer Pseudomonas Syringae as an artificial ice nucleus for weather modification[J]. Weather Modification, 1989, 21: 9～13.
[73] KRIEG. Bergry's Manual of Determination Bacteriology, 8th edition[M]. Williams ￡ Wilkins. Co.Baltimore. MD 21202, U.S.A. 1984, 140～476.
[74] BOCHANAN, R.E.Bergey's Manual of Systematic Bacteriology, 9th edition[M]. Williams ￡ Wilkins. Baltimore. MD 21202, U.S.A.1974, 217～340.
[75] 李宪利，高东升，陈广利，等．平原地区杏树花期推迟技术的研究[J]．落叶果树，1996，1：15～17．
[76] J.LAMAR ANDERSON, SCHUYLER, D.SEELEY. Bloom delay in deciduous fruits[J]. Hort. Rev., 1993, 15: 97～144.
[77] 郗荣庭．果树栽培学总论(第三版)[M]．北京：中国农业出版社，1997，303．
[78] LINDOW, S.E. The role of bacteria ice nucleation in frost injury to plant [J]. Ann. Rev. Phytopatbol. 1983, 21: 362～384.
[79] LINDOWS, S. E.Erwinia herbicola: a bacterial ice nucleus active in increasing frost damage to corn[J]. Phytopathology, 1978, 68: 523～527.
[80] MAKI L.R., GALYON E L, CHANG-CHIEN, M. et al. Ice nueleation induced by Pseudomonas syringae[J]. Appl. Mierobiol, 1974, 28: 456～460.
[81] SUN F Z, ZHU H, HE L Y, et al. Identification of Ice Nucleation Active Bacteria on Plants of China[J]. Progress in Natural Science, 1994, 4(3): 449～456.
[82] 孙福在，赵廷昌，牟丰盛，等．生防菌和药剂防除冰核细菌防御玉米霜冻危害研究[J]．自 然灾害学报，2003，12(4)：115～120．
[83] VALI G. Quantitative evaluation of experimental results on the heterogeneous freezing of supercooled liqnid[J]. AtmonSci. 1971, 28: 402～409.
[84] SCHAAD, W.N.Laboratory Guide for Identification of Plant Pathogenic Bacteria, 2nd edition[M]. St. Paul, Minnesota: The American Phytopathological society. 1988, 37～80.
[85] 任欣正．植物病原细菌的分类和鉴定[M]．北京：中国农业出版社，1999．
[86] 孙福在，朱红，何礼远，等．我国植物上冰核细菌种类鉴定[J]．自然科学进展，1994，4(4)：449～456．
[87] 刘招舰，丁爱云，张新建，等．山东省植物上冰核活性细菌种类与分布研究[J]．山东农业大学学报(自然科学版)，2002，33(3)：326～330．
[2] 罗心平，尼章光，王跃全，等．蛋黄果引种试种研究初报[J]．热带农业科技，2004，27(1)：47～48．
[3] 尼章光，罗心平，王跃全，等．值得开发的热带亚热带果树[J]．中国南方果树，2003，32(1)：25～26．
[4] 肖鸿修．中国热带南亚热带果树[M]．北京：中国农业出版社，1998．341～342．
[5] 冯玉香，何维勋．细菌冰核提高印度谷螟过冷却点的研究[J]．昆虫学报，1996，39(1)：53～57．
[6] 孙忠富．霜冻灾害与防御技术[M]．北京：中国农业出版社，2001．10～15．
[7] 唐广，蔡涤华，郑大玮．果树蔬菜霜冻与冻害的防御技术[M]．农业出版社，1993．30～57．
[8] 冯玉香，何维勋，崔松林．霜冻时冰晶在作物体内生长的试验研究[J]．中国农业气象，1991，12(4)：6～9．
[9] LEVITT, J.Freezing temperature responses of plants to environmental stress[M]. New York: Academic Press. 1979, 1: 267～291.
[10] PALTA, J.P., JEMSEM, et al. Cell membrane alterations following slow free thaw, injury and recovery[M]. Plant Cold Hardiness and Freezing Stress. New York: Academic Press, 1982, 2: 221～242.
[11] 何维勋，冯玉香，夏满强．解冻速率对农作物霜冻害的影响[J]．应用气象学报，1993，4(4)：440～445．
[12] 郭修武，傅望衡．葡萄根系抗寒性的研究[J]．园艺学报，1989，16(1)：17～22．
[13] 杨建民，李艳华，杨敏生，等．几个仁用杏品种抗寒性的比较[J]．中国农业科学，1999，32 (1)：46～50．
[14] 王飞，陈文登，高爱琴，等．杏品种一年生休眠枝、花、幼果抗寒的相关分析[J]．西北植物学报，1999，(4)：618～622．
[15] 彭伟秀，杨建民．杏花器官组织抗寒性研究[J]．果树学报，2002，19(2)：108～110．
[16] 白琳，白靳来．杏树花期、幼果期抗寒力调查[J]．北方园艺，1992，2(6)：69～70．
[17] 孙福在，朱红，何礼远．影响冰核细菌成冰活性的因素研究[J]．中国农业科学．1991，24(3)：57～64．
[18] 孙福在．我国生物冰核研究进展[J]．中国农业科学．1996，29(5)：62～68．
[19] OBATA, H., TAKINAMI, K, TANISHITA, J.T., et al. Identification of a new nucleation bacterium and its ice nucleation properties[J]. Agric Biol Chem, 1990, 54: 725～730.
[20] ORSER, C., STASKAWICZ, B.J., LOPER, J. et al. Cloning of genes involved in bacterial ice nucleation and fluorescent pigment production[J]. Agric Biol Chem, 1983, 12: 353～361.
[21] 高桥辛吉．INA细菌研究现状用化[J]．冷冻(日)，1987，62(718)：73～80．
[22] POOIEY, L., BROWN, T.A. Preparation of cell-free ice nuclear from Pseudomonas syringe[J]. Proc R Soc Lond Set B Biol Sci, 1990, 241: 112～115.
[23] 刘建华，陶毓汾．冰核细菌与玉米和大豆霜冻关系的研究[J]．中国农业气象，1990，11(2)：1～6．
[24] EDWARD, A.R., VANDENBUSSCHE, R.A., WICHMAN, H.A., et al. Unusual pattern of bacterial ice nucleation gene evolution[J]. Mol Biol Ecol, 1994, 11: 911～920.
[25] 杨建民，周怀军．果树霜冻害研究进展[J]．河北农业大学学报，2000，23(3)：54～58．
[26] 孙福在，赵廷昌．杏树上冰核细菌种类及冰核活性与杏花霜冻关系的研究[J]．中国农业科学，2000，(6)：50～58．
[27] LINDOW S E. The role of bacteria ice nucleation in frost injury to plant[J]. Ann. Rev. Phytopathol. 1983, 21: 326～384.
[28] ARAI, S. WATANABE, M.Freeze texturing of food materials by ice-nucleation with the bacterium Erwinia ananas[J]. Agric. boil. Chem., 1986, 50(1): 169～175.
[29] 杨建民，孟庆瑞．冰核细菌对杏花抗寒性的影响[J]．园艺学报，2002，29(1)：20～24．
[30] 韩伟秀，杨建民．冰核细菌对仁用杏花粉超微结构的影响[J]．园艺学报，2001，(5)：453～456．
[31] 孙福在，赵廷昌．冰核细菌生物学特性及其诱导植物霜冻机理与防霜应用[J]．生态学报．2003，23(2)：336～346．
[32] 张世光．PCR在快速检测冰核细菌及其冰核活性强度中的应用[A]．海峡两岸植物病理学术研讨会会刊[C]．中华植物病理学会，1997，4．
[33] PHELPS P, GIDDINGS TH, PHOCHODA M, et al. Release of cell-free ice nuclei by Erwinia herbicola[J]. Bactedol, 1986, 167: 496～502.
[34] MASANORI, KOIKE. Random Amplified Polymorpruz DNA Analysis of Japanese Isolates of verticilitun dahliae and Valbo-atrum[J]. Plant Disease, 1996, 80: 1224～1227.
[35] BASSI, A.S., DING, D.N., GLOOR, G.B., et al. Expression of a single chain antibodies(ScFVs) for c-myc oncoprotein in recombinant Escherichia coli membranes by using the ice-nucleation protein of Pseudomonas syringae[J]. Biotechnol. Prog., 2000, 16: 557～563.
[36] 唐朝荣．冰核细菌Erwinia ananas 110冰核基因克隆、序列分析和促冻杀虫基因工程菌构建[D]．中国农业科学院博士论文，2001．
[37] 唐朝荣，孙福在，赵廷昌，等．冰核细菌Erwinia ananas 110冰核基因克隆和序列分析[J]．微生物学通报，2002，29(4)：45～51．
[38] BIGG, E.K. The supercooling of water[J]. Proc Phys Soc B, 1953, 66: 688～694.
[39] ORSER C., STASKAWICZ, B.J., LOPER, J. et al. Cloning and expression of bacterial ice nucleation genes in Escherichia coli[J]. Bacterial. 1985, 164: 359-366.
[40] 姬广海，张世光．冰核活性细菌冰核基因多态性研究[J]．云南农业大学学报，1997，12(3)：153～157．
[41] WARREN, GJ., COROTTO, L., WOLBER, P.Conserved repeats in diverged ice nucleation structural genes from two species of Pseudomonas[J]. Nucl Acids Res, 1986, 20: 8047～8060.
[42] WOLBER, P., WARREN, G.J. Bacterial ice nucleation proteins[J]. TIBS, 1989, 14: 179.
[43] 任修海．冰核细菌的生物学及生物技术开发[J]．生物技术通报，1993，(6)：12～15．
[44] KOZLOFF, L.M., SCHOFIELD, et al. Ice nucleating activity of Psedomonas syringae and Erwinia herbicola[J]. Baceteriol., 1983, 153: 222～231.
[45] SCHNELL, R.C., VALI,G. Wodd-wide source of leaf-derived freezing nucIei[J]. Nature, 1973, 246: 212～213.
[46] WATANABE. M, ARAI.S. Bacterial ice nucleation activity and its application to freeze concentration of fresh food for modification of their properties[J]. Food Eng, 1994, 22: 453～473.
[47] LEEUNG, KWON MIKYUNG, SEONG KIYONG, et al. Development of a specific antibody for the detection of ice nucleation-active bacteria[J]. Plant Disease and Agriculture, 1999, 5(1): 27～33.
[48] TURNER, ARELLANOF, KOZLOFFM. Components of ice nucleation structures of bacteria[J]. Bacterial, 1991, 173: 6515～6527.
[49] 孙福在，朱红，何礼远，等．冰核细菌对光肩星天牛幼虫促冻杀虫研究[J]．林业科学研究，1997，10(1)：96～99．
[50] 张耀东，曹阳．冰核活性细菌对储粮害虫耐冻性的影响[J]．郑州粮食学院学报，1998，19(2)：1～5．
[51] 杨建民．INA细菌与杏树花器官霜冻害关系研究[D]．中国农业大学博士论文，2002，59-73．
[52] 王凤才．对2001年“三·二八”果树特大冻害的评估分析[J]．落叶果树，2002，3：11～14．
[53] 吕增仁．我国杏研究进展[J]．河北果树，1996，29(5)：62～67．
[54] 冯玉香．黄瓜霜冻与冰核细菌活性细菌的关系[J]．园艺学报，1990，17(3)：211～216．
[55] 何维勋，冯玉香，孙福在，等．防御霜灾新途径的研究[J]．灾害学，1990，(1)：14～19．
[56] 冯玉香，何维勋，夏满强．作物霜冻与低温强度及冰核菌密度的关系[J]．应用气象学报，1995，6(1)：90～91．
[57] 韦建福．我国生物冰核研究进展[A]．植物病虫害生物学研究进展一植物病虫害生物学国家重点实验室研究论文选[C]．北京：中国农业科技出版社，1995．
[58] ARNY D.C., LINDOW S E, UUPPER C D. Frost sensitivity of Zea may be increased by application of Pseudomnas Syingae[J]. Nature, 1976, 266: 282～284.
[59] LINDOW S E. Competitive exclusion of epiphytic bacteria by ice-mutants of Pseudomonas syringae[J]. Appl and Environ Microbiol, 1987, 53: 2520～2527.
[60] 朱红，孙福在，张永祥．冰核细菌对棉铃虫结冰温度影响的研究[J]．中国农业科学，1994，27(6)：23～27．
[61] 方中达．植病研究方法(第三版)[M]．北京：中国农业出版社，1998．
[62] 孙福在，赵廷昌，何维勋，等．应用生防菌和药剂防除冰核细菌控制玉米霜冻研究[M]．北京：中国农业科技出版社，2001．
[63] 张世光．云南冰核细菌的鉴定及防霜新技术研究[J]．云南农业大学学报，1995，10(2)：192～197．
[64] KIM, K.C., LEE, U., SONG, D.U. et al. Purification and characterization of ice nucleation-active bacteria[J]. Korean L Plant Pathol, 1996, 12(1): 99～108.
[65] WOLBE, P.K. Bacterial ice nucleation[J]. Adv. Microb. Physiol., 1993, 34: 203～207.
[66] WISNIEWSKIM., GLENN, D.M., FULLER, M.P.Use of a hydrophobic particle film as a barrier to extrinsic ice nucleation in tomato plants[J]. Hort Sci, 2002, 127(3): 358～364.
[67] 杨丽．冰核细菌活性的冰核基因克隆及其在大肠杆菌中表达[J]．中国农业科 学，1993，26(5)：88～89．
[68] LINDOW, S.E. Control of epiphytic ice nucleation-active bacteria for management of plant frost injury[M]. In: Biological Ice Nucleation and its Applications. Lee RE, Warren G and Gusta LV(eds). APS Press. 1996.
[69] GARETHE, WARREM. Bacteria Ice Nucleation: Molecular Biology and Application[J]. Biotechnology and Genetic Engineering Review, 1987, 5 (7): 88～89.
[70] LI, J.K., LEE, J.C. Bacterial ice nucleation and its potential application on the food industry. Trend Food Sci, 1995, 6: 259～263.
[71] 任修海．成冰核细菌的生物学及生物技术开发[J]．生物技术通报，1993，(6)：12～15．
[72] WARD P.J., DEMOTT P.J. Preliminary experimental valuation of SNOMAX snow inducer Pseudomonas Syringae as an artificial ice nucleus for weather modification[J]. Weather Modification, 1989, 21: 9～13.
[73] KRIEG. Bergry's Manual of Determination Bacteriology, 8th edition[M]. Williams ￡ Wilkins. Co.Baltimore. MD 21202, U.S.A. 1984, 140～476.
[74] BOCHANAN, R.E.Bergey's Manual of Systematic Bacteriology, 9th edition[M]. Williams ￡ Wilkins. Baltimore. MD 21202, U.S.A.1974, 217～340.
[75] 李宪利，高东升，陈广利，等．平原地区杏树花期推迟技术的研究[J]．落叶果树，1996，1：15～17．
[76] J.LAMAR ANDERSON, SCHUYLER, D.SEELEY. Bloom delay in deciduous fruits[J]. Hort. Rev., 1993, 15: 97～144.
[77] 郗荣庭．果树栽培学总论(第三版)[M]．北京：中国农业出版社，1997，303．
[78] LINDOW, S.E. The role of bacteria ice nucleation in frost injury to plant [J]. Ann. Rev. Phytopatbol. 1983, 21: 362～384.
[79] LINDOWS, S. E.Erwinia herbicola: a bacterial ice nucleus active in increasing frost damage to corn[J]. Phytopathology, 1978, 68: 523～527.
[80] MAKI L.R., GALYON E L, CHANG-CHIEN, M. et al. Ice nueleation induced by Pseudomonas syringae[J]. Appl. Mierobiol, 1974, 28: 456～460.
[81] SUN F Z, ZHU H, HE L Y, et al. Identification of Ice Nucleation Active Bacteria on Plants of China[J]. Progress in Natural Science, 1994, 4(3): 449～456.
[82] 孙福在，赵廷昌，牟丰盛，等．生防菌和药剂防除冰核细菌防御玉米霜冻危害研究[J]．自 然灾害学报，2003，12(4)：115～120．
[83] VALI G. Quantitative evaluation of experimental results on the heterogeneous freezing of supercooled liqnid[J]. AtmonSci. 1971, 28: 402～409.
[84] SCHAAD, W.N.Laboratory Guide for Identification of Plant Pathogenic Bacteria, 2nd edition[M]. St. Paul, Minnesota: The American Phytopathological society. 1988, 37～80.
[85] 任欣正．植物病原细菌的分类和鉴定[M]．北京：中国农业出版社，1999．
[86] 孙福在，朱红，何礼远，等．我国植物上冰核细菌种类鉴定[J]．自然科学进展，1994，4(4)：449～456．
[87] 刘招舰，丁爱云，张新建，等．山东省植物上冰核活性细菌种类与分布研究[J]．山东农业大学学报(自然科学版)，2002，33(3)：326～330．