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2025年8月15日 星期五
  2016, Vol. 24 Issue (12): 1925-1934    
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
巨大芽胞杆菌WY4的GFP标记及其在小白菜上的定殖
王珍1,曹翠玲1,库永丽1,徐国益1,林雁冰2,柳晓东1
1. 西北农林科技大学
2. 西北农林科技大学 生命科学学院
Bacillus megatherium WY4 Labeled by GFP and Its Colonization in Chinese Cabbage (Brassica chinensis)
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摘要 解磷菌(phosphate solubilizing bacteria, PSB)可以通过提高土壤有效磷含量而增加作物产量,目前已有许多解磷菌被分离并应用于农业生产中,但关于解磷菌在植物根际中的定殖情况仍缺乏系统性的研究。WY4为本实验室前期从小白菜(Brassica chinensis)根际分离得到的一株高效解磷菌,本研究利用绿色荧光蛋白(green fluorescent protein, GFP)标记技术研究了WY4在小白菜根际及土壤中的定殖规律。与原菌株相比,GFP标记对菌株WY4-GFP生长及解磷活性具有较小影响,同时在促进小白菜生长上WY4-GFP与WY4无显著性差异;WY4-GFP具有持久的定殖能力(接种21 d的自然土及30 d的小白菜根际土中,WY4-GFP的定殖数量分别为105和104 CFU/g左右),同时随时间的增加WY4-GFP在土壤中定殖数量逐渐减少;WY4-GFP在小白菜根冠及分生区大量定殖,在伸长区及侧根根毛处数量较少,同时表皮细胞间隙上也有较多的标记菌株。研究结果表明,WY4-GFP在小白菜根际及土壤中具有良好的定殖能力,这为后期深入研究解磷菌与植物间的关系提供了重要参考。
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王珍
曹翠玲
库永丽
徐国益
林雁冰
柳晓东
关键词 解磷细菌绿色荧光蛋白小白菜定殖    
Abstract:Phosphate solubilizing bacteria (PSB) can increase available phosphorous in soil and improve yields in several crop species like rice (Oryza sativa), maize (Zea mays), potato (Solanum tuberosum), sunflowers (Helianthus annuus) etc. In previous studies, many phosphate solubilizing bacteria had been isolated and widely used in agricultural production. However, the colonization of phosphate solubilizing bacteria in plant rhizosphere have not been completely characterized. Phosphate solubilizing bacteria WY4 is a bacterium isolated from the soil of Brassica chinensis rhizosphere soil. WY4 is a gram-positive bacterium which is tested by Gram staining and its cell morphology is rod shape. The sequence analysis of 16S rRNA gene in the NCBI database using a BlastN analysis showed 99% similarity to Bacillus megaterium strain. To evaluate colonization in soil and Chinese cabbage rhizosphere by this microorganism, generated WY4-GFP, a green fluorescent protein (GFP) labeled derivative of WY4, and inoculated the marked strain into Chinese cabbage seedlings. Compared with the original strain, GFP-labeled had a smaller influence on the growth and phosphate solubilizing activity. While, the results demonstrated that WY4-GFP could survive in the natural soil to achieve 105 CFU/g (wet soil) at 21 days; WY4-GFP increased the plant height, root length, leaf numbers, fresh weight and dry weight of Chinese cabbage plants. Phosphorus could promote root growth of Chinese cabbage, WY4 could increase content of available phosphorus as PSB, so the root system was developed after application of WY4: The root volume that reached 1.72 mL was significantly higher than water treatment (0.38 mL). After the treatments of WY4-GFP and WY4, the fresh weight of the plant reached 17.48 and 17.52 g/plant respectively. There was no significant difference between WY4 and WY4-GFP in growth promotion. The results further indicated that WY4-GFP flourishing colonizes in Chinese cabbage plants and rhizosphere soils, colonization of WY4-GFP analysis in nature soil or rhizosphere soils showed that the population of WY4-GFP decreased gradually as the extension of time. At 0~10 d, The number of WY4-GFP decreased very fast (from 13.32~3.47×106 CFU/g), but after 20~30 d incubation in Chinese cabbage rhizosphere soils, the number of WY4-GFP were basically stabled at 104 CFU/g. Furthermore, the results indicated that Bacillus megaterium WY4 displayed good colonization ability in the rhizosphere of Chinese cabbage. Plentiful WY4-GFP colonized in different growth period (5, 10 and 20 d) of the root tips and was detected in meristernatic zone. Meanwhile, less WY4-GFP appeared to the root elongation zone, lateral root hair and hypocotyls catheter, but numerous labeled bacteria existed in intercellular spaces of Chinese cabbage roots. Taken together, these results indicated that WY4-GFP invaded Chinese cabbage through the root tips or intercellular spaces, and they could be transported into the hypocotyls catheter by the vascular bundle system of roots. Preliminary study on the mechanism of phosphorus solubilizing bacteria promoting the growth of Chinese cabbage showed the good application prospects of PSB. The results provide important information for further study on the relationship between phosphate solubilizing bacteria and plant.
Key wordsPhosphate solubilizing bacteria (PSB)    Green fluorescent protein (GFP)    Chinese cabbage    Colonization
收稿日期: 2016-06-22      出版日期: 2016-11-07
ZTFLH:  Q939.95  
基金资助:陕西省科学技术研究发展计划项目
通讯作者: 曹翠玲     E-mail: cuilingcao@tom.com;caocuiling1@sina.com
引用本文:   
王珍 曹翠玲 库永丽 徐国益 林雁冰 柳晓东. 巨大芽胞杆菌WY4的GFP标记及其在小白菜上的定殖[J]. , 2016, 24(12): 1925-1934.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2016/V24/I12/1925
 
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