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1株桑叶内生促生细菌的筛选及对小麦幼苗的促生机制研究
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Abstract:
为获得植物内生促生菌资源，本文从桑树(Morus alba L.)叶片中分离筛选促生细菌，进行分类鉴定，检测其促生特性，探讨其对小麦(Triticum aestivum L.)的促生机制。结果显示，常规平板法从四个桑树品种叶片中分离出13株内生细菌，其中品种9703中分离的编号AG-2菌株表现出良好的促生效果。结合形态学和16S rRNA序列分析鉴定菌株AG-2为多黏类芽孢杆菌(Paenibacillus polymyxa)。平板定性法和高效液相色谱–串联质谱法(HPLC-MS/MS)分析表明菌株AG-2具有产有机酸、蛋白酶、纤维素酶、铁载体以及吲哚乙酸(IAA)、异戊烯基腺嘌呤(IP)和异戊烯基腺嘌呤核苷(IPA)的促生特性。室内幼苗浇灌试验结果表明，与对照组相比，菌株AG-2处理小麦幼苗12 d后，苗鲜重、根鲜重和单株总鲜重显著增加(P < 0.05)，增长率分别为12.16%、32.39%和24.68%；根冠比和根系活力变化不明显；叶片叶绿素含量和类胡萝卜素含量分别增加了180%和15.79% (P < 0.05)；叶片可溶性糖含量、脯氨酸含量和丙二醛(MDA)含量变化不明显，但可溶性蛋白含量增加显著(P < 0.05)；叶片超氧化物歧化酶(SOD)、苯丙氨酸解氨酶(PAL)和多酚氧化酶(PPO)活性显著增强(P < 0.05)，分别较对照组增加了113.14%、17.68%和68.24%。菌株AG-2能够有效促进小麦幼苗生长，具有开发成农业用微生物菌株的潜力。
In order to obtain endophytic bacteria to promote plants growth, strains were isolated and screened from mulberry leaves. Based on identification of the strain and analysis of the growth-promoting characteristics, the growth-promoting mechanism on wheat was explored. Results showed that 13 endophytic bacteria were isolated from the leaves of four mulberry varieties using conventional plate method, in which strain AG-2 from variety 9703 showed excellent growth-promoting effects. Through morphology and 16S rRNA sequence analysis, strain AG-2 was identified as Paenibacillus polymyxa. Plate qualitative analysis and high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) analysis showed that strain AG-2 had the promoting properties of producing organic acid, protease, cellulase, siderophore, as well as indole-3-acetic acid (IAA), isopentenyladenine (IP), and isopentenyladenine nucleoside (IPA). Compared with the control group, after 12 days of irrigating wheat seedlings in greenhouse, shoot fresh weight, root fresh weight and total fresh weight per plant significantly increased (P < 0.05), with growth rates of 12.16%, 32.39%, and 24.68%, respectively. Differences in root/shoot ratio and root vitality were not significant. Contents of chlorophyll and carotenoid in the leaves increased significantly (P < 0.05) by 180% and 15.79%, respectively. Changes in soluble sugar content, proline content, and malondialdehyde (MDA) content in leaves were not significant, but soluble protein content significantly increased (P < 0.05). In addition, activities of superoxide dismutase (SOD), phenylalanine ammonia lyase (PAL) and polyphenol oxidase (PPO) enhanced
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