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作者简介:

廖海枝(1997-),硕士研究生,主要从事果树生理与栽培研究,(E-mail)2362746801@qq.com。

通讯作者:

周开兵,博士,教授,主要从事果树生理与栽培研究,(E-mail)zkb@hainanu.edu.cn。

中图分类号:Q945.15

文献标识码:A

文章编号:1000-3142(2022)12-2138-09

DOI:10.11931/guihaia.gxzw202106008

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目录contents

    摘要

    为探讨叶面喷施钙镁肥对‘妃子笑’荔枝果肉苹果酸积累的影响,该文对‘妃子笑’荔枝树冠作喷布0.3%氯化钙(Ca)、0.3%氯化镁(Mg)及其二者混合(Ca+Mg)等水溶液处理,以树冠喷布清水为对照(CK),测定不同生长时期果肉水溶性钙和镁、苹果酸等含量及苹果酸代谢相关酶活性的动态变化,并作多元线性相关分析。结果表明:(1)苹果酸含量呈“L”型变化,Mg、Ca和Ca+Mg处理在果实发育前期促进苹果酸积累,Ca处理在后期促进苹果酸积累。(2)果肉水溶性钙含量总体呈上升趋势,水溶性镁含量大致呈“M”的动态变化趋势。(3)CK和Ca处理的苹果酸含量与NADP-ME活性、Ca+Mg处理的苹果酸含量与PEPC和NAD-MDH活性等均呈正相关,CK的苹果酸含量与PEPC活性、MS活性呈负相关。(4)水溶性钙抑制NAD-MDH、NADP-ME等活性,水溶性镁抑制NAD-MDH、MS等活性。综上认为,钙、镁叶面营养通过改变水溶性钙、镁等含量和苹果酸代谢途径不同关键酶活性而影响果肉苹果酸积累,其中Ca处理可能通过积累更多的苹果酸而抑制果肉呼吸作用,进而使果肉减少糖分损失,在生产中可作施肥技术应用。该研究结果为我国荔枝实际生产提供一定的理论参考和技术支持。

    Abstract

    In order to explore the effect of malic acid accumulation, spraying the foliar calcium and magnesium of ‘Feizixiao’ litchi, during the period of ‘Feizixiao’ litchi fruit development, 0.3% magnesium chloride (Mg), 0.3% calcium chloride (Ca), their mixture (Ca+Mg) and clean water (CK) were sprayed on leaves, and contents of water-soluble calcium, water-soluble magnesium and malic acid and the activities of malic acid metabolism-related enzymes in fruit flesh were measured. The multivariate linear correlation analysis were also performed. The results were as follows: (1) The content of malic acid showed a L-shaped trend, Mg, Ca and Ca+Mg treatments might promote the accumulation of malic acid in the early stage of fruit growth and development, while Ca treatment might promote the accumulation of malic acid in the late stage. (2) The content of water-soluble calcium in the flesh showed increasing trend, and the content of water-soluble magnesium showed the trend like “M”. (3) The content of malic acid was positively correlated with the activities of NADP-ME in CK and Ca, and PEPC and NAD-MDH in Ca+Mg, while the content of malic acid was negatively correlated with the activities of PEPC and MS in CK. (4) The water-soluble calcium inhibited the activities of NAD-MDH and NADP-ME, while water-soluble magnesium inhibited the activities of NADP-MDH and MS. In conclusion, foliar spraying of calcium and magnesium fertilizers can affect the contents of water-soluble calcium, magnesium and malic acid metabolism-related enzymes, and the linear correlation also changes. Different treatments can affect the accumulation of malic acid in pulp and cause the change of total acid content in pulp. Ca treatment may inhibit pulp respiration by accumulation of malic acid and then reducing the loss of sugar in pulp, which can be used as fertilizer technology in production. This results provide the theoretical reference and technical support for the actual production of litchi in China.

  • ‘妃子笑’荔枝(Litchi chinensis cv. Feizixiao)是我国荔枝主栽品种之一,是海南产区栽培面积最大的品种,具有较高的经济和社会效益(陈业光等,2008)。有机酸组分与含量是‘妃子笑’荔枝果实品质风味的重要组成因素(朱慧芹,2013)。‘妃子笑’荔枝果实的主要有机酸为苹果酸,为苹果酸型果实(周先艳等,2015),苹果酸含量是影响‘妃子笑’荔枝果实品质的重要影响因子。苹果酸是一种重要的初级代谢产物,在调节苹果渗透压、pH稳态、抗逆性和果实品质等方面起到关键作用(Zhang et al.,2020)。苹果酸作为‘妃子笑’荔枝果实的主要有机酸,对总酸含量有重要影响(乔方等,2012)。因此,研究荔枝苹果酸的积累特性,对丰富荔枝果实发育理论具有重要意义。

  • 近年来,国内外学者对果实有机酸代谢及其生理机制问题进行了研究,如糖酵解反应、三羧酸循环、糖异生作用等途径均存在着有机酸的踪迹(周先艳等,2015)。植物果实有机酸代谢过程极其复杂,其中有机酸代谢相关酶与有机酸含量密切相关(郭润姿等,2013)。苹果酸为植物果实重要有机酸之一,研究其代谢尤为重要,目前在不同植物有机酸代谢中,除烯醇式磷酸丙酮酸羧化酶(PEPC)均为关键酶外(Berüter,2004),在不同植物上曾报道过苹果酸脱氢酶(NAD-MDH)(Maldonado et al.,2004)、苹果酸酶(NADP-ME)(Crecelius et al.,2003)、苹果酸合成酶(MS)(Surendranathan &Nair,1976)等分别为其果实苹果酸代谢途径的相关酶。温清玉等(2012)报道,在荔枝中,PEPC是苹果酸合成的关键酶,NAD-MDH和NADP-ME也会影响苹果酸含量,但因品种不同而存在差异。此外,在不同果实上也存在相似情况。在‘蜂糖李’果实发育前期,苹果酸含量的变化由PEPC和NADP-ME协同调控,NAD-MDH作用不大; 与之不同的是,‘四月李’果实中引起苹果酸含量变化的关键酶是NAD-MDH与NADP-ME(王小红等,2018)。郭润姿等(2013)报道,苹果酸脱氢酶和苹果酸酶在黄冠梨果实发育中对苹果酸的产生与降解有重要作用。此外,温度、养分、品种遗传性等也会引起果实糖酸风味的变化(张红,2009)。刘洁云等(2021)报道,营养元素会对果实品质存在较大影响,如硒可以提高香蕉的果实品质。营养元素对果实酸度也有较大影响,氮、磷、钾、铜、铁等营养元素会影响果实酸度(陈发兴等,2005)。可见,苹果酸代谢仍处于探索阶段。因此,研究苹果酸代谢机制及如何调节苹果酸积累进而调节总酸含量是‘妃子笑’荔枝产业健康发展亟须解决的科学问题。

  • 本课题组前期研究发现施肥技术会影响矿质营养代谢而影响果实品质(苏阳等,2015b)。此外,还筛选出了能缓解‘妃子笑’荔枝果肉“退糖”现象(果面全红时果肉含糖量发生下降的现象)(苏阳等,2015a)的钙镁肥处理,其机制除了要关注果肉糖代谢变化问题外,果肉有机酸代谢是否发生变化的问题也不能忽视。鉴于此,本研究通过对‘妃子笑’荔枝树冠进行叶面喷施钙、镁肥处理,观测不同生长时期果肉水溶性钙和镁、苹果酸等含量及苹果酸代谢相关酶活性的动态变化,比较不同处理和对照间的差异,探讨叶面喷施钙、镁肥对果肉苹果酸积累的影响,以期探索调控果实酸含量的栽培措施提供理论依据,进而有效调控荔枝果实风味品质。

  • 1 材料与方法

  • 1.1 材料

  • 荔枝果实采摘于海南省临高县金牌农场五队荔枝园。选取营养状况良好、生长状况相近、无病虫害、株、行距6 m×7 m,冠幅约3 m×4 m的16年生‘妃子笑’荔枝果树20株。试验期间对20株果树采取一致的肥水管控、防病虫害管理措施。

  • 1.2 试验设计

  • 设置以下处理:(1)树冠叶面喷布0.3%氯化钙水溶液(Ca处理);(2)树冠叶面喷布0.3%氯化镁水溶液(Mg处理);(3)树冠叶面喷布0.3%氯化钙和0.3%氯化镁混合水溶液(Ca+Mg处理);(4)喷清水为对照(CK)。单株区组,重复5次。在每株样树的树冠中部外围四方选5个大小基本一致且生长中庸的果实进行挂牌标记,试验期间以这5个果的平均纵、横径为标准,在树冠中部外围选取对应大小的果实作为样果。每次处理前先取好果样30个,处理时间为谢花后35、42、50 d(上午900—1000),共处理3次,此后分别继续于谢花期后56、63、69、73 d取果样,共取样7次,果样就地放入液氮罐速冻,并储存于-80℃超低温冰箱中备用。

  • 1.3 方法

  • 苹果酸含量测定:参考胡志群等(2005)与王芮东等(2016)的方法并略有改动,将流动相换为0.1%磷酸二氢钠溶液,用磷酸调pH至2.8。

  • 苹果酸代谢途径相关酶活性测定:采用 Hirai和Ueno(1977)、罗安才等(2003)的方法制备酶液且测定酶活性。用酶标仪在450 nm波长下测定吸光度(OD值),通过标准曲线计算样品中烯醇式磷酸丙酮酸羧化酶(PEPC)、苹果酸脱氢酶(NAD-MDH)、苹果酸酶(NADP-ME)、苹果酸合成酶(MS)的酶活性。

  • 水溶性钙、镁含量测定:称取1 g左右荔枝果肉,连续烘干至恒重后加水研磨至匀浆,用去离子水震荡过夜后待测。采用火焰原子吸收法测定水溶性钙、水溶性镁含量(殷丽等,2013),使用仪器为NOVAA400P原子吸收分光光度计。

  • 1.4 数据统计分析

  • 利用SAS软件统计分析数据,采用ANOVA过程作方差分析和DUNCAN法作多重比较分析,用REG过程作多元线性相关性分析。

  • 2 结果与分析

  • 2.1 苹果酸含量变化

  • 如图1所示,在‘妃子笑’荔枝果实发育过程中,所有处理的苹果酸含量的变化趋势均呈现“L”型,花后50 d前急剧下降,后期趋于稳定。花后42 d,CK显著低于其余3个处理,而Mg处理又显著高于Ca处理; 花后50~69 d,所有处理间均无差异显著性; 花后73 d,Ca处理显著高于CK和Mg处理。由此可知,Ca、Mg和Ca+Mg处理只影响了前期苹果酸的积累且存在促进作用,后期仅Ca处理呈现促进趋势。

  • 图1 不同处理下苹果酸含量变化

  • Fig.1 Content changes of malic acid under different treatments

  • 2.2 苹果酸代谢途径相关酶活性变化

  • 2.2.1 磷酸烯醇式丙酮酸羧化酶(PEPC)

  • 如图2所示,CK的PEPC酶活性随果实发育进程历经3次“上升、下降”交替过程。Mg处理在35~42 d呈“平缓、上升”趋势,随后与CK趋势趋于一致; Ca处理则表现为“下降、上升、平缓、下降”的趋势; Ca+Mg处理呈“上升、下降、平缓、下降、上升、下降”的趋势。花后42 d,CK显著高于其余3个处理,Ca+Mg处理又显著高于Mg和Ca处理; 花后50 d,Ca处理显著最高; 花后56 d,Mg和Ca处理显著高于CK和Ca+Mg处理,CK又显著高于Ca+Mg处理; 花后63 d,Ca处理显著最高,CK又显著高于Mg和Ca+Mg处理; 花后69 d,CK显著高于Ca+Mg处理; 花后73 d,CK和Ca处理显著高于Mg和Ca+Mg处理。由此可知,Ca处理在花后50 d之后呈高于CK和其余处理趋势,Ca+Mg处理全程低于CK,Mg处理后期也具有低于CK的趋势,说明Ca处理呈提高PEPC活性的趋势,而Mg和Ca+Mg处理呈抑制PEPC活性的趋势。

  • 图2 不同处理下PEPC活性动态变化

  • Fig.2 Dynamic changes of PEPC activities under different treatments

  • 2.2.2 NAD-苹果酸脱氢酶(NAD-MDH)

  • 由图3可知,不同处理和CK的果肉NAD-MDH活性具有不同的动态变化趋势。CK呈“上升、平缓、下降、上升”趋势; Mg处理呈“下降、上升、平缓、上升、下降”趋势; Ca处理先无明显变化,随后历经两次“下降、上升”交替过程; Ca+Mg处理呈两次“下降、上升”交替趋势。由图3还可知,花后42 d,CK和Ca处理显著高于Mg和Ca+Mg处理; 在花后50~56 d,CK均显著最高,Mg处理又显著高于Ca+Mg处理,其中在50 d时为Mg处理显著高于Ca处理,在56 d时Mg处理显著低于Ca处理; 花后63 d,Mg处理显著高于Ca处理; 在69 d时Mg处理显著高于CK和其余处理; 花后73 d,Mg处理显著低于CK和其余处理,其余处理与CK无显著差异。由此可知,在63 d前CK持续高于其余处理,在63 d后Mg处理与CK趋势相反,说明Ca和Ca+Mg等处理均呈抑制NAD-MDH活性趋势,Mg处理则呈前抑后促的趋势。

  • 图3 不同处理下NAD-MDH活性动态变化

  • Fig.3 Dynamic changes of NAD-MDH activities under different treatments

  • 2.2.3 NADP-苹果酸酶(NADP-ME)

  • 由图4可知,不同处理和CK的果肉NADP-ME活性具有不同的动态变化趋势。CK呈“平缓、上升、下降、平缓、上升、平缓”的趋势; Mg处理呈“平缓、下降、上升、下降”趋势; Ca处理先无显著变化,随后呈“下降、平缓、下降”的趋势; Ca+Mg处理在35~50 d无明显变化,随后显著下降再上升。由图4还可知,花后42 d,Ca处理显著高于CK和其余处理; 花后50 d,CK显著最高,Mg处理又显著高于Ca处理; 花后63 d,Mg处理显著高于CK和其余处理,而CK和Ca处理又显著高于Ca+Mg处理; 花后69 d和73 d,CK酶活均为最高,均显著高于Ca处理,其中在69 d时Mg和Ca处理显著高于Ca+Mg处理,在73 d时Mg处理又显著高于Ca处理。由此可见,Ca、Mg和Ca+Mg处理均呈低于CK的趋势而呈抑制NADP-ME活性趋势。

  • 2.2.4 苹果酸合成酶(MS)

  • 由图5可知,不同处理和CK的果肉MS活性具有不同的动态变化趋势。在花后35~56 d,CK和Ca+Mg处理均呈现“下降、上升、下降”趋势,随后CK维持稳定趋势,Ca+Mg处理则持续降低至63 d后又上升; Ca处理初始并无显著变化,随后历经两次“下降、上升”交替过程后趋于稳定; Mg处理表现为“上升下降—上升下降”的趋势。由图5还可知,花后42 d,酶活由高往低顺序依次为Mg、Ca、CK、Ca+Mg处理,任意两者间均具有显著差异性; 花后50~63 d,CK均为显著最高; 在56 d和63 d时,Ca和Mg处理又均显著高于Ca+Mg处理,其中在63 d时Mg处理高于Ca处理; 花后69 d,Mg处理显著高于CK和其余处理,CK和Ca处理又显著高于Ca+Mg处理; 花后73 d,CK显著高于Mg和Ca+Mg处理。由此可知,Ca和Ca+Mg处理呈低于CK趋势,即呈抑制MS酶活性的趋势; Mg处理在42 d和69 d时表现提高酶活性的作用。

  • 图4 不同处理下NADP-ME活性动态变化

  • Fig.4 Dynamic changes of NADP-ME activities under different treatments

  • 图5 不同处理下MS活性动态变化

  • Fig.5 Dynamic changes of MS activities under different treatments

  • 2.3 水溶性钙、镁含量变化

  • 2.3.1 水溶性钙含量

  • 由图6可知,4个处理下花后63 d前的水溶性钙含量均呈现上升趋势,63 d后CK和Ca处理持续上升,Mg处理先上升至69 d再下降,Ca+Mg处理则维持稳定。由图6还可知,花后42 d,按Ca、Mg、CK、排序依次降低且任意两者间均显著; 花后50 d,Mg处理显著最高,Ca处理又显著高于Ca+Mg处理; 花后56 d,Mg处理仍为显著最高,CK又显著高于Ca和Ca+Mg处理; 花后63、69、73 d,Ca、Mg、Ca+Mg处理分别呈现显著最低、最高、最低。可见,Mg处理高于CK和其余处理; 而Ca+Mg处理则全程低于CK和Mg处理,说明Mg处理具有提高果肉水溶性钙含量的效果; 而Ca+Mg处理呈降低水溶性钙含量的趋势,Ca处理在73 d之后呈超越CK和其余处理的趋势。

  • 图6 不同处理下水溶性钙含量动态变化

  • Fig.6 Content changes of water-soluble calcium under different treatments

  • 2.3.2 水溶性镁含量

  • 由图7可知,从花后35 d开始至56 d,不同处理和CK的水溶性镁含量均先上升再下降; 在56~69 d,CK维持稳定,Ca+Mg处理先无明显变化后显著下降,Ca处理先下降再上升,Mg处理则呈“下降、上升、下降”趋势。由图7还可知,花后42 d,CK显著高于其余处理,Ca+Mg处理又显著低于Mg和Ca处理; 花后50 d,Mg处理显著最高,花后56 d,Ca处理均显著高于CK和Ca+Mg处理; 花后63 d,CK和Ca+Mg处理显著高于Mg和Ca处理; 花后73 d,Ca+Mg处理显著低于CK和Ca处理,Ca处理又显著高于CK。可见,Mg和Ca处理能提高果肉水溶性镁含量,Ca+Mg处理则呈降低水溶性镁含量的趋势。

  • 2.4 多元线性相关

  • 2.4.1 苹果酸含量与相关酶活性的多元线性相关

  • 对CK和不同处理的苹果酸含量与PEPC、NAD-MDH、NADP-ME、MS 4种酶活性作多元线性相关性分析,结果如表1和表2所示。由表1可知,CK和Ca处理的NADP-ME及Ca+Mg处理的PEPC、NAD-MDH等活性分别与苹果酸含量呈正相关; CK的PEPC、MS等活性与苹果酸含量则呈负相关。表明不同处理会改变酶活性与苹果酸含量的线性相关性,不同施肥处理能调节苹果酸的积累,并具有较为复杂的调节机制。由表2可知,除CK的苹果酸外,3个处理的苹果酸含量与其4种酶活性的复相关系数分别显著或极显著,这也说明苹果酸积累是这些酶共同作用的结果,任意一种酶活性改变均会引起苹果酸含量的改变,不同处理可能通过影响这些酶活性而影响苹果酸含量。

  • 图7 不同处理下水溶性镁含量动态变化

  • Fig.7 Content changes of water-soluble magnesium under different treatments

  • 表1 不同处理下苹果酸含量与相关酶活性的显著偏相关系数

  • Table1 Significant partial correlation coefficients between contents of malic acid and related enzyme activities under different treatments

  • 注: *表示显著性差异(P<0.05); **表示极显著性差异(P<0.01)。下同。

  • Note: *represents significant differences (P<0.05) ; **represents extremely significant differences (P<0.01) . The same below.

  • 2.4.2 水溶性钙、镁含量与相关酶活性的多元线性相关

  • 对所有处理的水溶性钙、镁含量分别与PEPC、NAD-MDH、NADP-ME、MS四种酶活性作多元线性相关性分析,结果如表2和表3所示。

  • 表2 不同处理下苹果酸、水溶性钙、水溶性镁等含量分别与相关酶活性的复相关系数

  • Table2 Multiple correlation coefficients between contents of malic acid, water-soluble calcium and water-soluble magnesium with related enzyme activities under different treatments

  • 表3 不同处理下水溶性钙、镁含量与相关酶的显著偏相关系数

  • Table3 Significant partial correlation coefficients between contents of water-soluble calcium and water-soluble magnesium with related enzymes under different treatments

  • 由表3可知,不同处理与CK水溶性钙、镁含量与相关酶活性的显著偏相关系数均为负相关,其中,CK的水溶性钙抑制NAD-MDH的酶活性,仅Mg处理的水溶性钙对NADP-ME的酶活无抑制作用; Ca+Mg、Mg处理的水溶性镁分别抑制NAD-MDH、MS活性。说明不同处理改变了水溶性钙、镁含量与4种酶活性的线性相关性,且不同的施肥处理通过调节苹果酸代谢相关酶活性而调节苹果酸的积累。由表2可知,3个处理和CK的水溶性镁含量与其4种相关酶活性的复相关系数均无显著性,而CK、Ca和Ca+Mg处理的水溶性钙含量与4种酶的复相关系数达显著或极显著水平。这说明叶面喷施钙、镁能改变果肉水溶性钙含量,使果肉水溶性钙含量与苹果酸代谢途径相关酶的活性产生线性相关性,进而调节果肉苹果酸的积累。

  • 3 讨论与结论

  • 钙存在多种形态,除水溶性钙外,还存在果胶酸钙、草酸钙、磷酸钙等(刘剑锋等,2004)。裴健翔(2019)研究表明,在苹果果实发育过程中,果实总钙和水溶性钙含量逐渐降低,果胶钙则先降后升,草酸钙含量逐渐升高,磷酸钙变化不明显,此外,采前钙处理可以增加果实中的总钙和水溶性钙含量(魏树伟和王少敏,2018)。本研究结果显示,经过烘干后4个处理的水溶性钙含量总体呈上升趋势,除受基因遗传影响外,也可能由于烘干后各种形式的钙转化为水溶性钙,因此总体表现为上升趋势。不同处理在花后63 d后的趋势表现出差异,63 d后CK和Ca处理持续上升,Mg和Ca+Mg处理则与之不同,这可能为Ca2+和Mg2+等离子间的复杂作用引起。本研究结果表明,单独喷施钙或镁营养分别促进果肉中水溶性钙、镁积累增多,Ca+Mg处理则会抑制水溶性钙、镁的积累。丁玉川等(2012)在甘蓝上报道钙、镁的吸收可能存在协同关系,本课题组前期研究中水溶性钙和镁存在相互增益效应(高丹等,2017),在猕猴桃叶片上Ca和Mg存在相互抑制吸收作用(刘科鹏,2013)。可见,水溶性钙、镁积累除了受遗传差异影响外,可能存在同时喷施钙、镁营养可能还会抑制植物体吸收钙、镁矿质元素。

  • 在果实发育过程中,通常果实成熟时,酸度会降低(张秀梅等,2007)。Wang等(2006)研究表明荔枝的苹果酸成熟前呈下降趋势。本研究结果表明,‘妃子笑’荔枝果肉苹果酸含量的变化趋势与前人研究相似,表现为急剧下降后逐渐趋于稳定; Ca、Mg和Ca+Mg处理前期呈促进影苹果酸积累的趋势,在果实生长发育后期仅Ca处理呈促进苹果酸积累的趋势,由于有机酸是呼吸代谢中间产物,其积累可能负反馈调节果肉呼吸作用(杨春宁等,2016),这可能导致Ca处理果肉呼吸代谢在后期较弱,从而积累糖分较多,即出现本课题组前期研究结果:Ca缓解果肉“退糖”现象(苏阳等,2015b)。对于本课题组前期研究结果Ca+Mg处理也具有缓解“退糖”现象,可能是Ca+Mg处理在果实生长发育后期促进其他有机酸的积累的缘故; Mg处理可能因为其与CK一样在果实生长发育后期未引起苹果酸和其他有机酸的积累改变。施肥改变了植物细胞体内细胞壁上的电荷变化与Mg2+等阳离子的竞争作用(李跃鹏等,2011),改变了介质中的H+和OH-的比例,从而改变了植物体内的pH值(郭悦等,2019),进而影响了有机酸含量。可见,叶面喷施钙镁肥会影响果肉中水溶性钙、镁的含量,Ca处理通过促进苹果酸的积累而促进总酸积累。

  • 苹果酸合成与丙酮酸羧化酶、苹果酸脱氢酶、苹果酸裂合酶、苹果酸酶有关(吴军林等,2014)。苹果酸合成酶(MS)是乙醛酸循环的关键酶(王程等,2011),乙醛酸循环为三羧酸循环的回补途径,与苹果酸的积累有重要作用。在枇杷果实的研究中,苹果酸的差异主要是NAD-MDH和NADP-ME的差异造成的,与PEPC也有关系(秦巧平等,2012)。马倩倩等(2017)研究表明,酸枣发育过程中苹果酸与NAD-MDH活性正相关,与NADP-ME活性负相关。李航等(2019)在樱桃果实上的研究也表明PEPC、NAD-MDH与苹果酸呈显著正相关,NADP-ME与苹果酸呈负相关。本研究显示,苹果酸含量与PEPC活性的相关关系因施肥处理不同而出现差异; CK和Ca处理的苹果酸含量与NADP-ME活性、Ca+Mg处理的苹果酸含量和NAD-MDH活性等呈正相关,CK的苹果酸含量与MS活性呈负相关。可见,本研究结果与前人研究结果并非完全一致,这可能与果实种类不同有关,说明不同施肥处理影响有机酸代谢具有不同的生化机理,作用于不同的靶标关键酶,进而引起苹果酸积累发生改变。本研究发现,叶面喷施钙、镁营养会影响苹果酸代谢相关酶活性,并且会改变苹果酸含量与苹果酸代谢相关酶活性的复相关关系。Ca2+和Mg2+能构成细胞渗透压,或起到活化酶,或成为酶和底物之间的桥接元素(Mengel et al.,2001),钙镁营养确实会影响酶活性及各种酶之间的联系,并且Mg和Ca处理在后期或有提高酶活的作用。离子间会存在协助作用(李联葆和王利平,2007),由于Ca2+具有稳定质膜结构的特殊功能,有助于质膜的选择性吸收,因此Ca2+对多种离子的吸收有协助作用(Inbal et al.,2010)。叶面钙镁营养改变了果肉水溶性钙、镁的积累,进而可能改变酶与其底物的结合特点,这可能是本研究钙镁营养影响酶活性及各种酶活性相关性的原因。

  • 综上所述,‘妃子笑’荔枝果实的苹果酸含量在果实发育过程中呈急剧减少后保持稳定的“L”型趋势,苹果酸积累差异受PEPC、NAD-MDH、NADP-ME和MS等4种关键酶共同调控,叶面喷施钙、镁通过影响4种关键酶活性及其与苹果酸含量的相关性而调节果肉苹果酸的积累,进而影响果肉的总酸含量和风味营养品质。叶面喷施钙肥通过促进苹果酸积累而抑制果肉呼吸作用,进而缓解妃子笑荔枝果肉成熟期“退糖”现象。关于叶面喷施钙、镁营养调节4种关键酶活性和苹果酸含量的详细生物学机制还有待深入研究。

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