一株人参内生产吲哚乙酸细菌的筛选及鉴定(1)
[摘要]研究以人参内生细菌为实验对象，采用Salkowski法筛选具有产吲哚乙酸(IAA)能力的菌株，同时采用Ashby培养基验证其固氮潜能，菌碟法及钼锑抗比色法测定其解磷能力，硅酸盐培养基初筛和火焰分光光度法复筛测定解钾能力，刃天青方法检测产生铁载体能力，α-酮丁酸法测定ACC脱氨酶活性;沙土法测定活性菌株对种子发芽的影响。结果从118株菌株中得到1株具有较高产IAA的菌株JJ5-2，其产IAA能力为10.2 mg·L-1;且具有解磷、解钾、固氮和产生铁载体特性;能显著促进人参种子根的生长;通过形态学、生理生化及16S rRNA基因序列分析鉴定菌株JJ5-2为苏云金芽孢杆菌Bacillus thuringiensis。
[关键词]内生细菌;促生特性;吲哚乙酸;人参
Screening and identification of indoleacetic acid producing
, 百拇医药 endophytic bacterium in Panax ginseng
JIANG Yun1， TIAN Lei1， CHEN Chang-qing2*， ZHANG Guan-jun1， LI Tong2， CHEN Jing-xiu1， WANG Xue2
(1.College of Life Science， Jilin Agriculture University， Changchun 130118， China;
2.College of Agronomy， Jilin Agriculture University， Changchun 130118， China)
[Abstract]Endophytic bacteria which was producing indoleacetic acid was screened from Panax ginseng by using the Salkowski method. The active strain was also tested for its ability of nitrogen fixation by using the Ashby agar plates， the PKV plates and quantitative analysis of Mo-Sb-Ascrobiology acid colorimetry was used to measure its ability of phosphate solubilization， for its ability of potassium solubilization the silicate medium and flame spectrophotometry was used， for its ability of producing siderophores the method detecting CAS was used， for its ability of producing ACC deaminase the Alpha ketone butyric acid method was applied. And the effect on promoting growth of seed by active strain was tested. The results showed that the indoleacetic acid producing strain of JJ5-2 was obtained from 118 endophytes， which the content of indoleacetic acid was 10.2 mg·L-1. The JJ5-2 strain also had characteristics of phosphate and potassium solubilization， nitrogen fixation， producing siderophores traits， and the promoting germination of ginseng seeds. The JJ5-2 strain was identified as Bacillus thuringiensis by analyzing morphology， physiological and biochemical properties and 16S rRNA gene sequences.
, 百拇医药
[Key words]endophytic bacteria; growth promoting; indoleacetic acid; Panax ginseng
doi：10.4268/cjcmm20150210
人参是我国最为重要的中药材之一，2012年又被卫生部批准为新资源食品，需求量逐年增加[1]。人参的种植对环境条件要求较高，在我国主要分布于吉林省长白山地区，现以人工栽培为主。为追求高产稳产，目前在人参种植的过程中广泛使用化学肥料和化学药剂，一定程度上降低了人参的品质，严重情况下甚至出现肥害和药害，导致化学制剂残留超标，同时也污染了种植区的生态环境。因此，发掘可促进人参生长的环境友好微生物，开发成微生物菌肥并加以应用，既可保障人参的产量和品质，又维护了种植区的生态环境，将促进人参产业健康可持续发展。目前有关植物促生菌报道主要包括产生长激素、固氮、解磷和解钾特性的微生物[2-4]，其中吲哚乙酸(IAA)是植物生长中所必需的生长调节剂，在植物生长过程中起着关键的作用，各个组织尤其是根部对IAA都比较敏感[5]。作为重要的微生物资源，植物内生和根际土壤细菌对于自身寄主具有较好的定殖和适应能力，可安全稳定的在寄主上进一步的应用。研究发现一些植物产IAA内生细菌能够显著促进植株的生长[6-7]，而关于人参内生产IAA细菌的研究未见报道。本研究拟从118株人参内生细菌群体中筛选具有产IAA活性兼具其他促生特性的菌株，鉴定其分类地位，以期发掘的促生菌株能够丰富植物促生菌资源，为人参微生物菌肥的开发与应用奠定基础。, 百拇医药(姜云 田磊 陈长卿 张冠军 李桐 陈京秀 王雪)
[关键词]内生细菌;促生特性;吲哚乙酸;人参
Screening and identification of indoleacetic acid producing
, 百拇医药 endophytic bacterium in Panax ginseng
JIANG Yun1， TIAN Lei1， CHEN Chang-qing2*， ZHANG Guan-jun1， LI Tong2， CHEN Jing-xiu1， WANG Xue2
(1.College of Life Science， Jilin Agriculture University， Changchun 130118， China;
2.College of Agronomy， Jilin Agriculture University， Changchun 130118， China)
[Abstract]Endophytic bacteria which was producing indoleacetic acid was screened from Panax ginseng by using the Salkowski method. The active strain was also tested for its ability of nitrogen fixation by using the Ashby agar plates， the PKV plates and quantitative analysis of Mo-Sb-Ascrobiology acid colorimetry was used to measure its ability of phosphate solubilization， for its ability of potassium solubilization the silicate medium and flame spectrophotometry was used， for its ability of producing siderophores the method detecting CAS was used， for its ability of producing ACC deaminase the Alpha ketone butyric acid method was applied. And the effect on promoting growth of seed by active strain was tested. The results showed that the indoleacetic acid producing strain of JJ5-2 was obtained from 118 endophytes， which the content of indoleacetic acid was 10.2 mg·L-1. The JJ5-2 strain also had characteristics of phosphate and potassium solubilization， nitrogen fixation， producing siderophores traits， and the promoting germination of ginseng seeds. The JJ5-2 strain was identified as Bacillus thuringiensis by analyzing morphology， physiological and biochemical properties and 16S rRNA gene sequences.
, 百拇医药
[Key words]endophytic bacteria; growth promoting; indoleacetic acid; Panax ginseng
doi：10.4268/cjcmm20150210
人参是我国最为重要的中药材之一，2012年又被卫生部批准为新资源食品，需求量逐年增加[1]。人参的种植对环境条件要求较高，在我国主要分布于吉林省长白山地区，现以人工栽培为主。为追求高产稳产，目前在人参种植的过程中广泛使用化学肥料和化学药剂，一定程度上降低了人参的品质，严重情况下甚至出现肥害和药害，导致化学制剂残留超标，同时也污染了种植区的生态环境。因此，发掘可促进人参生长的环境友好微生物，开发成微生物菌肥并加以应用，既可保障人参的产量和品质，又维护了种植区的生态环境，将促进人参产业健康可持续发展。目前有关植物促生菌报道主要包括产生长激素、固氮、解磷和解钾特性的微生物[2-4]，其中吲哚乙酸(IAA)是植物生长中所必需的生长调节剂，在植物生长过程中起着关键的作用，各个组织尤其是根部对IAA都比较敏感[5]。作为重要的微生物资源，植物内生和根际土壤细菌对于自身寄主具有较好的定殖和适应能力，可安全稳定的在寄主上进一步的应用。研究发现一些植物产IAA内生细菌能够显著促进植株的生长[6-7]，而关于人参内生产IAA细菌的研究未见报道。本研究拟从118株人参内生细菌群体中筛选具有产IAA活性兼具其他促生特性的菌株，鉴定其分类地位，以期发掘的促生菌株能够丰富植物促生菌资源，为人参微生物菌肥的开发与应用奠定基础。, 百拇医药(姜云 田磊 陈长卿 张冠军 李桐 陈京秀 王雪)