Bending Spore of Corn Leaf Spot Fungus REMI Albino Mutant Screening and Determination of Pathogenic Transformation

doi:10.13560/j.cnki.biotech.bull.1985.2014.12.018

Abstract

Abstract: To investigate bending spore corn leaf spot fungus pathogenic mechanism and genetic variation characteristics, this study using restriction enzymes mediated gene integration technology(REMI)of Curvularia lunata genetic transformation, received 109 stable mutant strains. PCR detection results showed that the plasmid pUC-JS has successfully imported mutant strains, into the son has both single copy and copy, albino strain single copy insert frequency was 40%. In addition, an albino corn bending spore bacteria leaf spot REMI contrast with wild strain of the mutant strains colony color, and also significantly weaker than wild on pathogenic strains. Plasmid save, sequencing and analysis of blast speculated that inserts may result in the loss FAD3 gene function.

Key words: Curvularia lunata REMI, Albino mutant, Pathogenic

Xia Shuchun,Wang Xuewu,Zhang Ruqin,Yan Honghai. Bending Spore of Corn Leaf Spot Fungus REMI Albino Mutant Screening and Determination of Pathogenic Transformation[J]. Biotechnology Bulletin, 2014, 0(12): 111-116.

References

[1] 吕国忠, 刘志恒, 何富刚, 等. 辽宁省爆发一种新病害-玉米弯孢菌叶斑病[J]. 沈阳农业大学学报, 1997, 28(1)：75-76.
[2] 陈捷, 鄢洪海, 高增贵, 等. 玉米弯孢菌叶斑病菌生理分化和寄主鉴定结果初报[J]. 植物病理学报, 2003, 4: 121-125.
[3] 郑战兵. 玉米弯孢霉叶斑病发生原因及综合防治[J]. 河南农业, 2012, 7: 32.
[4] 李永国, 关占良. 东北、华北春玉米生产主栽品种抗玉米弯孢叶斑病、丝黑穗病监测[J]. 中国植保导刊, 2012, 8: 41-44.
[5] 鄢洪海, 陈捷, 刘萍, 等. 玉米弯孢叶斑病菌白化菌株初报[J]. 菌物系统, 2002, 21(4)：604-606.
[6] Tanaka J, Shiotnai H, Yamamoto M, et al. Insertional mutagenesis and cloning of genes required for biosynthesis of the bots-specific AKtoxin in the Japanese pear Pathotype of Alternaria alternate[J]. Mol Plant-Microbe Interact, 1999, 12: 691-702.
[7] 张建萍, 朱凯, 杨爽, 等. 稗草生防潜力菌 Helminthosporium gramineum Rabehn f. sp. echinochloae 的原生质体制备和再生[J]. 浙江农业学报, 2010, 22(1)：14-19.
[8] Akamatsu H, Itoh Y, Kodama M, et al. AAL-toxin-deficient mutants of Alternaria alternata tomato Pathotype by restriction enzyme-mediated integration[J]. Phytopathology, 1997, 87: 967-972.
[9] George MLC, Nelson RJ, Zeigler RS, et al. Rapid population analysis of Magnaporthe grisea by using rep-PCR and endogenous repetitive DNA sequences[J]. Phytopathology, 1998, 88(3)：223-229.
[10] 李坡, 谷守芹, 刘丽华, 等. 玉米大斑病菌 REMI 体系的建立及突变体分析[J]. 中国农业科学, 2011, 44(4)：716-722.
[11] 张建萍, 朱凯, 余柳青. 产蛇孢菌素 A 禾长蠕孢 REMI 突变体的制备及其筛选[J]. 中国生物防治学报, 2013, 29(3)： 424-430.
[12] Sambrook J, Fritsch EF, Maniatis T. Molecular cloning a laboratory manual[M]. 2nd. New York(USA)：Cold Spring Harbor Laboratory Press, 1989.
[13] 李铁民, 孟晋. 限制性内切酶介导的整合技术[J]. 辽宁大学学报: 自然科学版, 2005, 32(4)：325-327.
[14] Lu S, Lynghholm L, Tagged G, et al. Tagging mutations at the Toxl locus of Cochliobolus heterostrophus by restriction enzyme-mediated integration[J]. Proc Natl Acad Sci, 1994, 91: 12649-12653.
[15] Shi Z, Christian D, Leung H. Enhanced transformation in Magnaporthe grisea by restriction enzyme mediated integration of plasmid DNA[J]. Phytopathology, 1995, 85(3)：329-333.
[16] Dye JM, Chapital DC, Cary JW, et al. Chilling-Sensitive post-transcriptional regulation of a plant fatty acid desaturase expressed in yeast[J]. Biophys Tes Commun, 2001, 282: 1019-1025.
[17] Inoue L, Ohara T, Namiki F, et al. Isolation of pathogenicity mutants of Fusarium oxysporum f. sp. melonis by insertional mutagenesi[s J]. Journal of General Plant Pathology, 2001, 67(3)： 191-199.
[18] 代玉华, 刘训言, 孟庆伟, 等. 低温胁迫对类囊体膜脂代谢的影响[J]. 植物学通报, 2004, 21(4)：506-511.
[19] Horiguchi G, Fuse T, Kawakami N, et al. Temperature-dependent translational regulation of the ER omega-3 fatty acid desaturase gene in wheat root tips[J]. Plant J, 2000, 24: 805-813.
[20] Zhou X, Xu S, Liu L, et al. Degradation of cyanide by Trichoderma mutants constructed by restriction enzyme mediated integration (REMI)[J]. Bioresource Technology, 2007, 98(15)：2958-2962.