Screening for microorganisms from aromatic plants is an important way to produce natural fragrances using biotechnology. Endophytic bacteria were isolated from five different strains of Cinnamomum longepaniculatum (Gamble) N. Chao leaves with various oil production rates and 1,8-cineole content. Using tissue block, grinding, and neutralizer auxiliary methods with lysogeny broth culture medium (LB), tryptose soya agar medium (TSA), and lecithin Tween 80 nutrient agar medium, endophytic bacteria were isolated and grown. Gas Chromatography-Mass Spectrometer (GC-MS) were used as quantitative analysis tool to screen for high α-Terpineol -yield strains and the high-yield strains were further mutated by UV. After removing the duplicate strains according to the morphological characteristics of the colony, 176 strains of endophytic bacteria in the leaves of Cinnamomum longepaniculatum (Gamble) N. Chao were obtained. These strains were divided into 4 phyla, 13 genera, and 34 species. The Bacillus was the first dominant class. Among the 34 strains of endophytic bacteria, the optimal mutagenicity of the strain T3-M-39 with high α-Terpineol production was obtained using a 30 W UV lamp with 30 cm irradiation for 2 minutes, which had a mortality rate of 74.1%. The production of α-Terpineol was enhanced with the mutant strain, reaching 1.45 mg/L, which was increased by 34.9% compared with that of the wild strain. In addition, this mutant strain was also able to maintain its stable genetic characteristics. This finding provided the basis for the natural α-Terpineol production using biotechnology.
[1] 罗中杰, 李维一, 魏琴, 等.宜宾油樟的现状及未来[J].四川师范大学学报(自然科学版), 2001,24(3):317-319.
[2] 尹礼国, 凌跃, 杜永华, 等.宜宾油樟营养器官精油主成分分析[J].江苏农业科学, 2014,42(11):348-350;355.
[3] 胡文杰, 许樟润, 李冠喜.基于响应面法对油樟籽油超声波提取工艺的优化[J].中国粮油学报, 2017,32(2):109-115.
[4] CHRISTINA K, NURHAYAT T, GERHARD B. α-Terpineol, a natural monoterpene: A review of its biological properties[J]. Open Chemistry, 2018,16(1):349-361.
[5] DANIELE N G, JANARA S C, MARLANGE A O, et al. α-Terpineol reduces cancer pain via modulation of oxidative stress and inhibition of iNOS[J]. Biomedicine & Pharmacotherapy, 2018,105:652-661.
[6] POLYANNA D S N, DOUGLAS S D C, VALDEANIA G D S, et al. Antidiarrheal activity of α-terpineol in mice[J]. Biomedicine & Pharmacotherapy, 2019,110:631-640.
[7] 陈绮琪,黄贵玲,刘文森, 等.碳硅复合固体酸催化剂的制备及其催化性能研究[J].肇庆学院学报, 2019,40(2):45-50;55.
[8] 牟大庆.二步法生产药用松油醇工艺技术改进及效果分析[J].生物质化学工程, 2010,44(1):27-30.
[9] 曹松,安兰芝,卢祥, 等.D型大孔磺酸树脂催化松节油水合一步合成松油醇[J].离子交换与吸附, 2015,31(5):455-463.
[10] 梁鸣,陈敏,蔡春平, 等.松油醇的分析及其生产工艺改进的研究[J].色谱, 2002,20(6):577-581.
[11] 田红玉,陈海涛,孙宝国.食品香料香精发展趋势[J].食品科学技术学报, 2018,36(2):1-11.
[12] CUI J L, WANG C L, GUO S X, et al. Stimulation of dragon's blood accumulation in Dracaena cambodiana via fungal inoculation [J]. Fitoterapia, 2013,87(1): 31-36.
[13] 严宽, 陈放, 魏琴, 等.内生真菌对油樟挥发油积累及生理生化特性的影响[J].生物技术通报, 2017,33(3):138-143.
[14] 周婧, 杨琦, 李钢, 等.红树属植物内生真菌多样性及其代谢产物研究进展[J].林业科学, 2019,55(1):92-102.
[15] 王涛, 游玲, 魏琴, 等.油樟内生细菌的多样性及抑制植物病原菌初步研究[J].西北林学院学报, 2009,24(2):97-100.
[16] 王涛, 游玲, 崔晓龙, 等.油樟内生芽孢细菌的系统发育多样性[J].微生物学报, 2009, 49(5):574-580.
[17] 方珍娟, 张晓霞, 马立安.植物内生菌研究进展[J].长江大学学报(自科版), 2018, 15(10):41-45.
[18] LI L, LI Z W, YIN Z Q, et al. Antibacterial activity of leaf essential oil and its constituents from Cinnamomum longepaniculatum [J]. International Journal of Clinical & Experimental Medicine, 2014,7(7):1 721-1 727.
[19] 魏琴, 周宇科, 周黎军, 等.油樟油抑制细菌生长的活性试验[J].热带农业科学, 2009,29(1):5-7.
[20] 刘晓丹,王俊玲,王霞.树舌灵芝复合诱变选育漆酶高产菌株[J].北方园艺, 2019(14):124-129.
[21] 田蕊,萨如拉,段晓霞, 等.产γ-氨基丁酸乳酸菌的筛选鉴定及其紫外诱变研究[J].食品工业科技, 2018,39(17):128-132.
[22] 孙会忠,宋鹏,杜君, 等.一株硅酸盐细菌的鉴定及其紫外线诱变育种[J].基因组学与应用生物学, 2018,37(3):1 225-1 230.
[23] 辛羚,俞苓,齐凤兰, 等.天然香精香料与生物技术[J].食品科技, 2004,25(11):49-51;54.
[24] 陈泽斌,夏振远,雷丽萍,等.烟草可培养内生细菌的分离及多样性分析[J].微生物学通报, 2011,38(9):1 347-1 354.
[25] 王玉美.汽爆辅助提取喜树碱及产喜树碱内生菌的筛选[D].北京:中国科学院大学,2011.
[26] 张庭静,蒋瑾,徐淑霞, 等.一株产梓醇的地黄内生细菌的鉴定[J].浙江农业科学, 2011(2):395-399.
[27] 陈奕涵,钱悦,侯永泰, 等.复合诱变选育大分子量透明质酸高产菌株[J].中国酿造, 2012,31(9):98-101.
[28] 张建新,张吨,王海磊,等.产β-mannanase内生菌的诱变育种和酶学特性研究[J].现代食品科技, 2012,28(10):1 331-1 335;1 410.
[29] 徐婉如.秦艽内生真菌的诱变育种及其发酵条件优化[D].西安:西北大学,2008.
[30] 张建新,赵杰,陈泽田, 等.产β-甘露聚糖酶内生菌的诱变育种及产酶条件优化[J].基因组学与应用生物学, 2014,33(4):815-821.
