• ISSN 1008-505X
  • CN 11-3996/S

氮磷钾配施对北苍术产量、品质及其相关酶基因表达的影响

Effects of combination of nitrogen, phosphorus and potassium on the yield, quality and the related enzyme gene expression of Atractylodes chinensis

  • 摘要:
    目的 研究氮磷钾配施对北苍术的产量、苍术素含量及乙酰辅酶A羧化酶(ACC)活性和基因表达量的影响,探讨氮磷钾配施对北苍术药材产量增加、质量形成的影响及其机制,对提高北苍术药材产量及质量有重要的现实意义。
    方法 以一年生北苍术为试验材料,采用 L9(34) 正交设计,设置10个不同氮、磷、钾用量处理的田间试验,在营养生长期、花期、初果期、盛果期及枯萎期测定北苍术的地上部分、根茎和须根鲜重;采用高效液相色谱法,测定北苍术中苍术素含量;采用双抗体夹心法(ELISA),测定北苍术根茎中乙酰辅酶A羧化酶(ACC)的活性;采用实时荧光定量PCR法(qRT-PCR),测定北苍术关键酶ACC基因的相对表达量;利用Spearman相关性分析,探讨苍术素合成的分子调控机制。
    结果 不同氮磷钾肥料配施均能够促进一年生北苍术地上部分以及地下根茎、须根鲜重的增加,其中T3 (N1P3K3)和T6 (N2P3K1)施肥方案最佳,地上部分鲜重在盛果期分别为对照组的1.52和1.47倍,地下根茎鲜重、须根鲜重在枯萎期分别为对照组的2.08和2.09倍、1.30和1.41倍。T3、T6施肥方案北苍术有效成分苍术素含量在枯萎期较对照组分别提升33%和61%,且根据不同生育期内苍术素的动态积累规律,推测北苍术枯萎期至次年出苗前是苍术素积累的最佳时期。通过相关分析表明,苍术素含量与ACC活性和ACC基因表达量之间的相关系数分别为0.66和0.583,ACC活性与ACC基因表达量相关系数达0.839,均达到极显著正相关(P < 0.01)。低、中氮水平下高磷处理提高了关键酶ACC基因的表达量,促进ACC活性提升,并进一步增加苍术素的积累,而高氮、高钾抑制了ACC基因的表达,进而降低苍术素的积累。
    结论 高磷有利于北苍术中 ACC 基因的表达及酶活性的提升,促进苍术素的合成和积累,有利于北苍术的提质增效。北苍术的最佳施肥方案为:氮肥90 kg/hm2、磷肥225 kg/hm2、钾肥315 kg/hm2 或氮肥180 kg/hm2、磷肥225 kg/hm2、钾肥105 kg/hm2

     

    Abstract:
    Objectives The suitable combination of nitrogen (N), phosphorus (P) and potassium (K) fertilizers was studied from the yield, atrctylodin content and the activity and gene expression of acetyl coenzyme A carboxylase (ACC) of A. chinensis, so to propose a scientific fertilizer combination for improving the productivity and quality of A. chinensis.
    Methods A field experiment including 10 combined applications of N, P and K fertilizers was set up following a L9(34) design, with one-year-old Atractylodes chinensis as experimental material. The fresh weight of aboveground part, rhizome, and fibrous root of A. chinensis were weighed at vegetative growth, flowering, early fruiting, full fruiting and withering stages. The plant samples were collected at the same time for determination of atrctylodin content by HPLC method, the ACC activity in rhizome by double antibody ELISA method, the relative expression level of ACC gene using real-time PCR. Spearman correlation analysis was used to determine the molecular regulation mechanism of atrctylodin synthesis.
    Results All the NPK combined applications promoted the growth of aboveground part, rhizome and fibrous root of the crop. The most significant effects were observed in T3 (N1P3K3) and T6 (N2P3K1), the fresh weight of aboveground part were respectively 1.52 and 1.47 times that from N0P0K0 at full fruiting period, the fresh weight of underground rhizome were 2.08 and 2.09 times of that from N0P0K0 at withering period, and the fresh weight of fibrous roots were 1.30 and 1.41 times of that from N0P0K0 at withering period. Compared with N0P0K0, T3 and T6 increased atrctylodin by 33% and 61% at withering period. Moreover, according to the dynamic accumulation characteristics of atrctylodin in different growth stages, it was speculated that the most important period for atrctylodin accumulation was the withering period to the next year before emergence. According to the results of correlation analysis, the correlation coefficients between atrctylodin content and ACC enzyme activity, ACC gene expression were 0.66 and 0.583, respectively. The correlation coefficient between ACC enzyme activity and ACC gene expression was 0.839, which reached extremely significant positive correlation (P<0.01). Treatment of high P with low and medium N rates improved the gene expression of ACC and enhanced the ACC activity thereby increased the accumulation of atrctylodin. While high level of N and K inhibited the gene expression of ACC, thereby decreased the accumulation of atrctylodin.
    Conclusions High level of P fertilizer (225 kg/hm2), combined with appropriate level of N (90 kg/hm2) and K fertilizer (315 kg/hm2) or P (225 kg/hm2), N (180 kg/hm2) and K fertilizer (105 kg/hm2) are beneficial to the yield of A. chinensis, which will stimulate the gene expression, enhance ACC activity, and promote the synthesis and accumulation of atrctylodin.

     

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