Patil Chandrashekhar, Suryawanshi Rahul, Koli Sunil, Patil Satish
School of Life Sciences, North Maharashtra University, Jalgaon, 425001, Maharashtra, India; Avram and Stella Goldstein-Goren Department of Biotechnology Engineering, Ben-Gurion University of the Negev, Beer Sheva 84105, Israel.
School of Life Sciences, North Maharashtra University, Jalgaon, 425001, Maharashtra, India.
J Microbiol Methods. 2016 Dec;131:102-104. doi: 10.1016/j.mimet.2016.10.009. Epub 2016 Oct 18.
Aminocyclopropane-1-carboxylate deaminase (ACCD) producing microorganisms support plant growth under a variety of biotic and abiotic stress conditions such as drought, soil salinity, flooding, heavy metal pollution and phyto-pathogen attack. Available screening methods for ACCD give idea only about its primary microbial ACCD activity than the actual potential. In the present investigation, we have simply improved screening method by incorporating pH indicator dyes (phenol red and bromothymol blue) in ACC containing medium. This modification is based on the basic principle that ACCD action releases ammonia which can be detected by color change and zone around the bacterial colony. High color intensity and zone around the colony indicates most potent producer, colony showing only a color change indicates moderate potential and no change in colony color indicates least efficiency. Enzymatic bioassays as well as root elongation studies revealed that ACC-deaminase activity of Pseudomonas aeruginosa, Stenotrophomonas maltophilia and Bacillus subtilis clearly corresponds to their growth on dye incorporated ACC medium. This method could be used to complement the existing screening methods and to speed up the targeted isolation of agriculturally important microorganisms.
产生1-氨基环丙烷-1-羧酸脱氨酶(ACCD)的微生物在多种生物和非生物胁迫条件下,如干旱、土壤盐分、洪水、重金属污染和植物病原体攻击,都能促进植物生长。现有的ACCD筛选方法只能了解其主要的微生物ACCD活性,而不是实际潜力。在本研究中,我们通过在含ACC的培养基中加入pH指示染料(酚红和溴百里酚蓝),简单地改进了筛选方法。这种改进基于这样一个基本原理:ACCD的作用会释放氨,氨可以通过颜色变化和菌落周围的区域检测到。菌落周围颜色强度高且有区域表明是最有效的生产者,仅显示颜色变化的菌落表明潜力中等,菌落颜色无变化表明效率最低。酶活性测定以及根伸长研究表明,铜绿假单胞菌、嗜麦芽窄食单胞菌和枯草芽孢杆菌的ACC脱氨酶活性明显与其在含染料的ACC培养基上的生长情况相对应。该方法可用于补充现有的筛选方法,并加速农业重要微生物的靶向分离。