• ISSN 1008-505X
  • CN 11-3996/S

产黄青霉菌对不同难溶性磷酸盐的溶解能力及作用机制

Mechanism and dissolve capacity of Penicillium chrysogenum to different insoluble phosphates

  • 摘要:
    研究目的 探究了产黄青霉菌(Penicillium chrysogenum)对不同难溶性磷酸盐(磷酸铁、磷酸铝和磷酸三钙)的溶解效率及机理。试验方法供试产黄青霉菌株为本研究筛选的高效解磷真菌菌株。试验采用蒙金娜液体培养基,分别加入磷酸铁(FePO4·2H2O,Fe-P)、磷酸铝(AlPO4,Al-P)和磷酸三钙Ca3(PO4)2,Ca-P作为难溶性磷源,接种产黄青霉菌孢子悬液培养。在培养1、3、5天后,过滤培养液,测定滤液pH、溶磷量、有机酸含量及酶活性,采用红外光谱和扫描电镜–能谱分析Fe-P、Al-P和Ca-P处理滤渣的矿物官能团和形貌变化。
    结果 青霉菌对难溶性磷酸盐的溶解量随着培养时间的延长而增加。在培养5天后,Fe-P、Al-P和Ca-P的溶磷量分别为153.8、215.9和569.4 mg/L,对Ca-P的溶解量最高。红外光谱图显示,Fe-P、Al-P和Ca-P的PO43−振动吸收峰强度发生了不同程度的减弱。随着培养时间的延长,培养液pH均呈下降趋势,在培养5天后,Fe-P、Al-P和Ca-P处理的培养液pH由培养初期的6.5分别下降至2.0、2.3和5.0。对于不同难溶性磷酸盐,青霉菌分泌的丙酮酸脱氢酶、柠檬酸合成酶活性以及有机酸含量不同,与Al-P和Ca-P处理相比,Fe-P处理显著增强了青霉菌中丙酮酸脱氢酶和柠檬酸合成酶活性,草酸和柠檬酸分泌量分别为1086.6和806.4 mg/L,而在Al-P和Ca-P处理中,分泌的有机酸主要是草酸,分泌量分别为261.1和201.3 mg/L。草酸对Fe-P、Al-P和Ca-P的释放率分别为14.8%、32.4%和39.6%,分别比柠檬酸高出37、3和2倍。通过扫描电镜发现,草酸可与Ca-P螯合形成草酸钙(CaC2O4)。
    结论 产黄青霉菌对不同难溶性磷酸盐的释放能力表现为Ca-P>Al-P>Fe-P。与Al-P和Ca-P相比,Fe-P能够提高产黄青霉菌丙酮酸脱氢酶和柠檬酸合成酶的活性,从而促进产黄青霉菌分泌更多的草酸和柠檬酸。虽然草酸溶解难溶性磷酸盐的能力强于柠檬酸,但是与Fe-P相比,Ca-P抑制了产黄青霉菌分泌草酸的能力。因此,利用产黄青霉菌活化土壤难溶性磷酸盐时,可通过添加Fe-P诱导提高其分泌柠檬酸和草酸的能力。

     

    Abstract:
    Objectives Penicillium chrysogenum (P. chrysogenum) is a typical phosphate solubilizing fungi used in microbial strains. We studied the capacity and mechanism of P. chrysogenum in dissolving P from ferric, aluminum and tricalcium phosphate (Fe-P, Al-P and Ca-P).
    Methods The tested P. chrysogenum was selected in our laboratory, and was isolated into Fe-P, Al-P and Ca-P suspension, respectively. After incubation of 1, 3, and 5 days, the suspension was filtered using a no-P filter paper, the filtrate was used for determination of soluble P, organic acids concentration, and enzyme activities. The filter residue were used for observation of morphology of phosphates and fungi mycelium using attenuated total reflectance infrared spectroscopy (ATR-IR) and scanning electron microscopy-energy dispersive spectrometer (SEM-EDS).
    Results The dissolve amount of P from the insoluble phosphates increased with the elongation of incubation days. P. chrysogenum dissolved more P from Ca-P (P<0.05) than from Al-P and Fe-P. After five days of incubation, the P content in Fe-P, Al-P and Ca-P was 153.8, 215.9, and 569.4 mg/L, respectively; and the filtrate pH of the Fe-P, Al-P, and Ca-P treatments decreased to 2.0, 2.3, and 5.0, respectively. The Fe-P filtrate had (P<0.05) higher pyruvate dehydrogenase and citrate synthase activities than the Ca-P and Al-P filtrates. The oxalic acid and citric acid concentration in the Fe-P filtrate was 1086.6 and 806.4 mg/L, respectively. While only oxalic acid was detected in Al-P and Ca-P filtrates, with concentration of 261.1 and 201.3 mg/L, respectively. The ATR-IR spectra showed that the vibration absorption peaks of PO43− in the Fe-P, Al-P, and Ca-P treatments became significantly weaker, indicating the release of P from the tested phosphates. The SEM-EDS analysis showed that the oxalic acid was chelated with Ca-P and formed calcium oxalate (CaC2O4). Compared with citric acid, oxalic acid was more efficient in the release of P from Fe-P, Al-P and Ca-P, with release rates of 14.8%, 32.4% and 39.6%, which was 37, 3 and 2 times higher than citric acid, respectively.
    Conclusions The capacity of P. chrysogenum in dissolving P from different insoluble phosphates is in the order of Ca-P>​Al-P>Fe-P. When exposure to Fe-P, P. chrysogenum enhances the activity of pyruvate dehydrogenase and citrate synthasesecretes to secrete more oxalic acid and citric acid. Although oxalic acid is more capable of dissolving insoluble phosphates than citric acid, its chelating with carbonates may offset its capacity. So addition of ferric phosphate might help increase the efficiency of P. chrysogenum to dissolve insoluble phosphates via the secretion of citric and oxalic acid in soil.

     

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