海洋细菌解淀粉芽胞杆菌SH-27在大豆体内的定殖动态及促生防病作用

doi:10.16409/j.cnki.2095-039x.2019.02.004

摘要/Abstract

摘要： 为明确海洋细菌SH-27在大豆体内的定殖动态及其促生长作用和对大豆疫病的防治效果，本研究采用抗利福平标记法和平板对峙生长法，筛选对利福平标记稳定且对大豆疫霉菌具有较好抑菌作用的标记菌株SH-27^Rif，培养10代后的标记菌株SH-27^Rif能够保持稳定，对大豆疫霉抑制率为56.92%。分别采用灌根和涂叶法研究其在大豆体内的定殖动态，灌根与涂叶法均可使标记菌株SH-27^Rif在大豆体内定殖，时间达31 d以上；灌根处理定殖量呈先升后降趋势，定殖量根 > 茎 > 叶，处理后第21 d根部定殖量达到最大（6.6×10⁵ cfu/g）；涂叶处理第1 d大豆叶片定殖量达到最大（6.3×10⁵ cfu/g），随后呈迅速下降趋势；定殖量叶>茎，根部未检测到标记菌株SH-27^Rif。盆栽促生试验结果表明，菌株SH-27发酵液灌根处理第15 d，处理组株高、根长、茎粗、鲜重、干重、叶绿素含量、根系活力等指标均显著高于对照组。盆栽防病试验结果表明：菌株SH-27发酵液灌根处理能显著降低大豆疫病的病情指数，对大豆疫病3、5、7和9 d的防效分别为83.44%、66.34%、57.18%和52.85%。以上研究结果表明海洋细菌SH-27是防治大豆疫病的潜在生防菌株，具有良好的开发和应用价值。

关键词: 海洋细菌, 大豆疫病, 定殖, 解淀粉芽胞杆菌

Abstract: Marine bacterium strain SH-27 was applied to soybean plants to investigate the colonization dynamics, growth promotion effect and the biological control effect on Phytophthora root rot disease. Mutant strainSH-27^Rif with stable marker of antibiotic rifampicin and inhibitory effect against Phytophthora sojae were screened by using the rifampin-resistant mutants and dual culture method. Colonizing dynamics of the strain SH-27^Rif in root, stem and leaf of soybean were investigated by treating with root irrigating and leaf daubing. Then the effects of growth promotion and controlling disease on soybean seedlings were studied by pot tests. The results indicated that the resistance marker of the mutant strain SH-27^Rif was remained stable after 10 generations of cultivation. The inhibition rates of strain SH-27^Rif and original strain SH-27 against P. sojae were 56.92% and 57.58%, respectively. The strain SH-27^Rif could colonize for 31 days or more in soybean seedlings by root irrigating and leaf daubing. After root irrigating, the colonization quantity of strain SH-27^Rif trended to increase firstly and then decrease. The highest quantity in roots was 6.6×10⁵ cfu/g at 21 d after inoculation. Bacteria quantities in different parts were root > stem > leaf. After leaf daubing, the highest colonization quantity in leave was 6.3×10⁵ cfu/g at 1 d after inoculation and then decreased quickly. Bacteria quantities in different parts were leaf>stem and strain SH-27^Rif wasn't detected in root. Pot tests demonstrated that the height, root length, stem diameter, fresh weight, dry weight, chlorophyll and root activity of soybean seedlings treated with SH-27 fermentation broth were significantly higher than those of the control groups. When challenging the pathogen Phytophthora sojae, soybean seedlings treated by SH-27 showed decreased disease index significantly and the control efficacies were 83.44%, 66.34%, 57.18% and 52.85% at 3 d, 5 d,7 d and 9 d, respectively. In conclusion, the results indicated that marine bacterium strain SH-27 was an excellent candidate for developing a biocontrol agent to control Phytophthora root rot of soybean.

Key words: marine bacteria, soybean Phytophthora root rot, colonization, Bacillus amylolique faciens

中图分类号:

S476

林巧玲, 卢乃会, 何红, 袁越. 海洋细菌解淀粉芽胞杆菌SH-27在大豆体内的定殖动态及促生防病作用[J]. 中国生物防治学报, 2019, 35(2): 240-246.

LIN Qiaoling, LU Naihui, HE Hong, YUAN Yue. Colonization Dynamics and Growth Promotion Effect of Marine Bacterium Strain SH-27 in Soybean and Its Biological Control Effect on Phytophthora Root Rot Disease[J]. journal1, 2019, 35(2): 240-246.

参考文献

[1] Wrather J A, Anderson T R, Arsyad D M, et al. Soybean disease loss estimates for the top ten soybean producing countries in 1998[J]. Canadian Journal of Plant Pathology, 2001, 23:115-121.
[2] Schmitthenner A F, Hobe M, Bhat R G. Phytophthora sojae races in Ohio over a 10 year interval[J]. Plant Disease, 1994, 78:269-276.
[3] Stewart S, Abeysekara N, Robertson A E. Pathotype and genetic shifts in a population of Phytophthora sojae under soybean cultivarrotation[J]. Plant Disease, 2014, 98:614-624.
[4] 付红梅, 李淼. 大豆疫霉拮抗菌株的筛选与鉴定[J]. 安徽农业科学, 2011, 39(19):11482-11484, 11495.
[5] 付思娅, 陈双林, 闫淑珍. 植物内生细菌在辣椒体内的定殖动态及对辣椒疫病的防治效果[J]. 中国生物防治学报, 2013, 29(4):561-568.
[6] Lugtenberg B, Kamilova F. Plant-growth-promoting rhizobacteria[J]. Annual Review of Microbiology, 2009, 63:541-556.
[7] Kamilova F, Validov S, Azarova T, et al. Enrichment for enhanced competitive plant root tip colonizers selects for a new class of biocontrol bacteria[J]. Environmental Microbiology, 2005, 7(11):1809-1817.
[8] 何红, 邱思鑫, 蔡学清, 等. 辣椒内生细菌BS-1和BS-2在植物体内的定殖及鉴定[J]. 微生物学报, 2004, 44(1):13-18.
[9] 胡伟, 赵兰凤, 张亮, 等. 香蕉枯萎病生防菌AF11的鉴定及其定殖研究[J]. 中国生物防治学报, 2012, 28(3):387-393.
[10] Thomas L, Ana Z, Tatiana K. Aromatic compounds degradation plays a role in colonization of Arabidopsis thaliana and Acacia caven by Cupriavidus pinatubonensis JMP134[J]. Antonie van Leeuwenhoek International Journal of General and Molecular Microbiology, 2012, 101(4):713-723.
[11] 高毓晗, 李世东, 郭荣君. sfp基因转化增强了Bacillus subtilis 168的定殖能力和对黄瓜茎内枯萎病菌的抑制作用[J]. 中国生物防治学报, 2016, 32(1):76-85.
[12] 刘海洋, 姚举, 张仁福, 等. 甲基营养型芽胞杆菌AL7对棉花黄萎病的盆栽防治效果及定殖能力[J]. 中国生物防治学报, 2017, 33(3):378-384.
[13] 黄勤知, 卢乃会, 何红, 等. 红树内生细菌AiL3在大豆体内的定殖与促生作用研究[J]. 大豆科学, 2014, 33(2):223-227.
[14] 杨光红, 高智谋, 曹舜, 等. 大豆疫病拮抗细菌的筛选及生防作用[J]. 安徽农业大学学报, 2012, 39(5):677-681.
[15] 杨定祥, 林巧玲, 卢乃会, 等. 拮抗辣椒疫霉菌海洋细菌菌株SH-27的筛选鉴定及其防病促生作用[J]. 微生物学通报, 2018, 45(1):54-63.
[16] 汪腾, 段雅婕, 刘兵团, 等. 两株香蕉枯萎病拮抗菌在香蕉体内的定殖[J]. 基因组学与应用生物学, 2011, 30(3):342-350.
[17] 韩祥东, 花美娜, 冯永君, 等. 拮抗大豆疫霉菌植物内生细菌的筛选与鉴定[J]. 大豆科学, 2012, 31(1):85-91.
[18] 陈申宽, 闫任沛, 王秋荣, 等. 大豆疫病分级标准与危害性研究[J]. 大豆通报, 2002(5):7.
[19] 秦娟娟, 闫淑珍, 刘佳. 植物内生细菌固体菌剂对辣椒的促生和防病作用[J]. 植物保护学报, 2010, 37(4):325-329.
[20] Compant S, Clement C, Sessitsch A. Plant growth-promoting bacteria in the rhizo-and endosphere of plants:their role, colonization, mechanisms involved and prospects for utilization[J]. Soil Biology and Biochemistry, 2010, 42(5):669-678.
[21] 蔡学清, 何红, 胡方平. 双抗标记法测定枯草芽胞杆菌BS-2和BS-1在辣椒体内的定殖动态[J]. 福建农林大学学报(自然科学版), 2003, 32(1):41-45.
[22] 王君, 马海林, 丁延芹, 等. 4株根际促生菌对核桃生长及生理特性的影响[J]. 山东农业科学, 2014, 46(4):77-79.
[23] 柳凤, 潘朝勃, 何红, 等. 海洋细菌对辣椒疫霉和辣椒疫病的抑制作用[J]. 中国生物防治, 2008, 24(4):379-381.
[24] 张兴锋, 柳凤, 何红, 等. 红树内生细菌CⅢ-1菌株鉴定及其胞外抗菌蛋白性质[J]. 微生物学通报, 2010, 37(2):222-227.
[25] 何培青, 王红梅, 沈继红, 等. 南极芽胞杆菌Bacillus sp.107生长特性及抑制植物病原真菌的研究[J]. 海洋科学进展, 2011, 29(2):179-185.
[26] Tareq F S, Hasan M C, Lee H S, et al. Gageopeptins A and B, new inhibitors of zoospore motility of the phytopathogen Phytophthora capsici from a marine-derived bacterium Bacillus sp. 109GGC020[J]. Bioorganic and Medicinal Chemistry Letters, 2015, 25(16):3325-3329.
[27] Mouchka M E, Hewson I, Harvell C D. Coral associated bacterial assemblages:Current knowledge and the potential for climate-driven impacts[J]. Integrative and Comparative Biology, 2010, 50(4):662-674.