湖南农业大学学报 ( 自然科学版 ) 2016, 42(3):312 316. DOI:10.13331/j.cnki.jhau.2016.03.017 Journal of Hunan Agricultural University (Natural Sciences) http://xb.ijournal.cn 荠菜提取液对烟草青枯病菌的抑制作用 1,2 1,2 1,2 1,2* (1. 410128 2. 410128) 摘要 63.9% 28.9% 关键 词 中图分类号 S435.72 文献标志码 A 文章编号 1007 1032(2016)03 0312 05 Inhibition activity of Capsella extracts against Ralstonia solanacearum Feng Youtao 1,2, Xie Yan 1,2, Zhang Ya 1,2, Liao Xiaolan 1,2* (1.College of Plant Protection, Hunan Agricultural University, Changsha 410128, China; 2. Hunan Provincial Key Laboratory for Biology and Control of Plant Diseases and Insect Pests, Changsha 410128, China) Abstract: Water, ethyl acetate, anhydrous ethanol and petroleum ether, acetone, methanol were used to extract Capsella powder, the inhibitory effects of 6 kinds of solvent extracts on Ralstonia solanacearum were investigated through indoor virulence test, the results showed that only ethyl acetate and water extracts from Capsella had antimicrobial activity on Ralstonia solanacearum, the of ethyl acetate extract reached 63.9%, and antibacterial rate of water extract was 28.9%. The influences of water and ethyl acetate extracts on morphology, growth curve,utilization rate of sugar and protein content of Ralstonia solanacearum were investigated by transmission electron microscopy (SEM), wheat flange dividing turbidimetry, anthrone colorimetry, coomassie brilliant blue method.electron microscopy showed that the structures of cell membrane and cell wall were changed and the bacteria cracked in Ralstonia solanacearum treated with water extract and ethyl acetate extract from Capsella of. Ethyl acetate extract and water extract from Capsella can make Ralstonia solanacearum logarithmic phase lag, reduce sugar utilization, and increase protein content in Ralstonia solanacearum. Keywords: Capsella extracts; Ralstonia solanacearum; inhibition (Capsella ) [1] Tarle [2] (Staphylococcus) (Escherichia coli ) [3] (Bacillus subtilis) (Staphylococcus aureus Rosenbach) (salmonella) Saman Soleimanpour [4] Woo Jin Choi [5] 收稿日期 :2016 03 10 修回日期 :2016 04 06 基金项目 ( ) (201303025) 作者简介 (1990 ) 971820275@qq.com * liaoxl88@126.com
42 3 313 (Ralstonia solanacearum) [6] [7] [8 9] [10] [11] 71.30% [12] 99% [13] 簕 1 材料与方法 1.1 材料 (Ralstonia solanacearum) 3 (NA) [14] ph7.0 7.2 1.2 方法 1.2.1 荠菜活性物质的提取 [15] 20 ml 4 20 ml 4 1.2.2 烟草青枯病菌菌悬液制备及抑菌活性测定 NA 28 48 h 3 10 8 cfu/ml [16] 1.2.3 荠菜活性提取液对病原菌细胞形态影响的观察 NA 1 ml 2 5 mm 140 r/min 28 24 h 400 µl 8 h 1.2.4 荠菜提取液对烟草青枯病菌生长曲线 糖利用率 蛋白质含量的影响的测定 [17] [18] [19] 200 µl 1.3 数据处理 DPS Excel 2010 2 结果与分析 2.1 荠菜不同溶剂提取液的抑菌效果 ( 1) 40 mm 60% 18 mm 28.9%
314 ( ) http://xb.ijournal.cn 2016 6 1 2 3 4 5 6 7 8 Fig.1 图 1 荠菜不同溶剂提取液的抑菌效果 Inhibitory effects of different solvent extracts from Capsella on Ralstonia solanacearum 2.2 荠菜无菌水和乙酸乙酯提取液对烟草青枯病菌细胞形态的影响 2 A ( 2 B) 2 C 2 D A B C D 图 2 烟草青枯病菌的扫描电镜观察结果 Fig.2 SEM observation of Ralstonia solannacearum
42 3 315 2.3 荠菜提取液对烟草青枯病菌生长曲线 糖利用率和蛋白质含量的影响 3 CK1 CK2 CK1 CK2 CK1 CK2 CK1 CK2 CK1 CK2 OD600 nm 2.5 2.0 CK1 CK2 水提 酯提 1.5 1.0 0.5 0.0 10 15 20 25 30 35 图 3 荠菜乙酸乙酯和水提液处理的烟草青枯病菌生长曲线 Fig.3 The growth curves of Ralstonia solannacearum treated by 4 4 CK1 CK2 /(mg ml 1 ) 25 20 15 10 5 0 CK1 CK2 水提 酯提 10 20 30 40 50 图 4 荠菜乙酸乙酯和水提液处理的烟草青枯病菌的含糖量 Fig.4 The sugar contents of Ralstonia solannacearum treated by CK1 CK2 /(mg ml 1 ) 1000 CK1 CK2 800 水提 酯提 600 400 200 0 2 4 6 8 图 5 荠菜乙酸乙酯和水提液处理的烟草青枯病菌的蛋白质含量 Fig.5 The protein contents of Ralstonia solannacearum treated by 3 讨论 6 参考文献 : [1] [J] 2003(3) 44 [2] Tarle Djurdjca Dvorzak et al Capsella bursapastoris(l.) medikus-investigation of antibiotic effect saponin and flavonoid substances[j] Fann Glas 1988 44(4)
316 ( ) http://xb.ijournal.cn 2016 6 111 116 [3] [J] 2010(4) 146 148 [4] Saman Soleimanpour Fereshteh Sadat Sedighinia Akbar Safipour Afshar et a1 Synergistic antibacterial activity of Capsella bursapastoris and Glycyrrhiza glabra against oral pathogens[j] Jundishapur J Microbiol 2013 6(8) 1 6 [5] Woo Jin Choi Seong Keun Kim Hee Kuk Park et a1. Anti-inflammatory and anti-superbacterial properties of sulforaphane from shepherd s purse [J] Korean J Physiol Pharmacol 2014(18) 33 39 [6] Hayward A C Biology and epidemiology of bacterial wilt caused by Pseudomonas solanacearum[j] Phytopathol 1991 29 65 87 [7] 1 [J] ( ) 2011 37(6) 637 640 [8] [J] 2013 33(4) 84 87 [9] [D]. 2012 [10] [J] 2009(3) 62 64 [11] [J] ( ) 2011 26(2) 284 287 [12] [J] 2015(5) 120 124 [13] 52 [J] 2011 31(6) 15 18 [14] [M] 2007 [15] [J] 2015(8) 49 53 [16] [J] 2012 37(1) 204 206 [17] [J] 2010 40(6) 601 608 [18] D [J]. 2016 19(1) 36 39 [19] [J] 2011 24(6) 53 55 责任编辑 : 罗慧敏英文编辑 : 罗维