不同矮化中间砧对华红苹果果实中总酚及组分含量的影响

李青山1,2,高 源2,王 昆2,孙思邈2,路 翔1,2,李连文2,冯建荣1*,王大江2*

1石河子大学农学院·特色果树栽培生理与种质资源利用兵团重点实验室,新疆石河子 832000;2中国农业科学院果树研究所·农业农村部园艺作物种质资源利用重点实验室,辽宁兴城 125100)

摘 要:【目的】探讨6个不同矮化中间砧对华红苹果果实中总酚及组分动态含量的影响,为苹果矮化中间砧的选择应用和优质苹果生产提供应用依据。【方法】以山荆子为基砧,CG24、MD001、SH3、SH38、Mac9和CX5为中间砧,嫁接华红为试验材料,山荆子直接嫁接华红为对照;在果实生长发育的7个时期,采用超高效液相色谱法及质谱联用技术测定多酚组分及含量、Folin-Ciocalteus法测定总酚含量。【结果】华红果皮中总酚含量在所有时期均高于果肉,且随着果实的发育逐渐降低,但在发育过程中出现最小值的时期不同:果肉在成熟期最低,果皮8月29日最低;果皮、果肉多酚组分含量随着果实发育和成熟逐渐降低或在果实发育阶段先降低,随着果实成熟增高,不同矮化中间砧组合对果皮、果肉多酚组分的动态变化有不同程度的影响。在成熟期,SH38组合、CG24 组合和SH3组合提高了华红果皮总酚含量,MD001组合和SH38组合提高了华红果肉总酚含量;Mac9组合和SH38组合显著提高华红花青苷含量,进而促进果实着色;SH38组合和MD001组合对果皮、果肉多酚组分的含量提升较多。【结论】SH38组合可考虑作为提高华红苹果功能性栽培的首选中间砧。

关键词:苹果;华红;矮化中间砧;总酚;多酚组分

中国是苹果种植大国,国家统计局资料显示,2020 年产量达4 406.60 万t[1],苹果产业在我国农业中的地位举足轻重,不仅在脱贫攻坚和美丽乡村建设中发挥作用,也对我们的生活健康大有益处。苹果富含酚类化合物,酚类物质是强抗氧化剂,对人体具有延缓衰老,抗菌消炎等保健功效[2-4]。苹果中晚熟品种华红,是由中国农业科学院果树研究所以金冠为母本、惠为父本杂交选育的红果品种,表现为果个大、外观美、风味浓、抗褐化等特性[5],然而华红果实中起抗褐化作用的酚类组分并不明确,产业中矮化中间砧对其多酚组分和含量的影响鲜有报道。研究华红果实发育过程中果皮和果肉中总酚和多酚组分的动态变化,以及不同矮化中间砧对其总酚和多酚组分的影响,探索其抗褐化的可能原因及作为功能果皮开发具有重要的意义。果实多酚物质的组成和含量不仅由品种本身特性决定,也受各种栽培条件的影响,不同砧木不仅可以增加果实产量[6-7],提升可溶性固形物含量[8]和糖酸比[9],增强树体对矿质营养的吸收[10],促进幼树提早开花[11],提高树体抗寒性[12],还可以提高果皮色素和多酚物质含量。矮化中间砧M26 组合(烟富6/M26/平邑甜茶)的果皮和果肉的总酚含量显著高于对照(烟富6/平邑甜茶)[13],520a 和101-14MG 砧木可明显提高矢富罗莎葡萄果肉中的酚类化合物的含量[14]。不同砧木影响了红亚历山大葡萄酚类物质含量,显著促进葡萄果实着色[15]。矮化密植栽培是当前新建果园主流的苹果种植模式,但在我国北方寒冷地区,由于自根砧不耐寒,采用矮化中间砧是实现寒冷地区苹果矮化密植的主要途径[16-18]。因此,笔者以探寻不同矮化中间砧对其酚类的影响和华红苹果果肉抗褐化的可能原因为出发点,通过6 种矮化中间砧对华红苹果果皮、果肉总酚及多酚组分和含量的分析,研究不同矮化中间砧对华红苹果总酚及多酚组分和含量的动态影响,探究提升华红苹果果实功能性作用的最适矮化中间砧,旨在为今后苹果抗褐化品种的选育和栽培提供理论依据。

1 材料和方法

1.1 试验材料

全部供试矮化中间砧6个(表1),2010年8月采用芽接方法在基砧山荆子上嫁接6 个矮化中间砧,2011年8月采用芽接方法在矮化中间砧上嫁接苹果品种华红。并采用相同方法在基砧山荆子上嫁接华红作为对照。2012年4月定植于辽宁省葫芦岛市韩家沟果树农场苹果矮化中间砧示范园,田间常规管理,中间砧的长度为25~30 cm。

表1 供试矮化中间砧及其选育单位
Table 1 Dwarfing interstocks and breeding units

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从2020 年6 月9 日开始到10 月8 日,每隔20 d采样1次,共取样7次。6月9日为果实膨大期初期,10月8日为果实成熟期。不同矮化中间砧组合和对照组华红苹果均采用相同的套袋管理,每个处理及对照选取树体长势一致的3 株树,于树体外围和中部采集生长发育一致的果实10 个,采摘后去除果核,削掉果皮,果肉切块,果皮和果肉液氮冷冻后放入-80 ℃冰箱保存待测。每株树的果实为1个重复,每个样品设置3次重复。

1.2 苹果总酚含量的测定

总酚含量的测定采用Folin-Ciocalteus法[19-20],用没食子酸作标准曲线。7 个组合、7 个时期果皮、果肉切碎,用组织捣碎机匀浆,取匀浆5 g,用80 mL蒸馏水洗入100 mL 容量瓶中,至100 ℃沸水浴30 min,冷却后定容至100 mL,过滤,吸取1.0 mL滤液,加入1.0 mL FoLin-CiocaLteus试剂溶液,用蒸馏水定容至10 mL 混匀,静置,室温显色,用紫外分光光度计TU-1800SPC(北京普析)在765 nm测定吸光度,查阅标准曲线,根据稀释倍数,计算样品总酚含量。

1.3 苹果多酚组分的提取纯化和测定

华红果实中的多酚组分的测定方法参照李静等[21]的方法,略有改动。

取出-80 ℃冰箱保存的样品,进行冷冻研磨,分别称取冷冻研磨后的果皮和果肉样品10 g,用80%甲醇分2 次超声提取,每次20 mL,合并提取液定容至50 mL,吸取10 mL 提取液在旋转蒸发仪上40 ℃减压旋蒸除去甲醇。依次用5 mL 甲醇活化固相萃取小柱,倒入样液,用5 mL 蒸馏水分2 次冲洗固相萃取小柱,弃除冲洗液,再用4 mL 甲醇分2 次洗脱固相萃取小柱,收集洗脱液,甲醇定容至5 mL,吸取1 mL提取液在旋转蒸发仪上40 ℃减压旋蒸除去甲醇,甲醇定容到5 mL,过0.22 μm有机相滤膜,待分析。

1.4 统计分析

研究采用Microsoft Excel 2016 软件处理数据,采用SPSS 26.0 软件统计分析,采用单因素ANOVA的LSD Duncan 比较处理间的差异显著性(p <0.05),采用Omicshare 平台做趋势分析,采用Origin 2019b软件绘图。

2 结果与分析

2.1 华红果实总酚含量的动态变化

选取对照(华红/山荆子)进行总酚含量的动态分析,由图1 可知,在果皮和果肉中,总酚含量均随着果实发育逐渐降低,在7 个发育时期果皮中总酚含量明显高于果肉;果皮、果肉总酚在果实发育前期显著下降,在果实发育中后期无显著性差异;总酚含量(w,后同)随着果实发育逐渐降低,出现最小值的时期不同,果肉在8月29日达最低,为409.24 mg·kg-1,9 月18 日略有上升,果实成熟(10 月8 日)下降至最小值,为350.31 mg·kg-1。在果皮中,总酚含量先迅速降低,然后缓慢升高,在8月29日最低,为1 143.48 mg·kg-1,成熟期含量上升为1 273.33 mg·kg-1;在成熟期果皮总酚含量是果肉的363.5%。

图1 华红果实总酚含量的动态变化
Fig.1 Dynamic changes of total phenolics in the fruit of Huahong apple

不同小写字母表示经Duncan 检验差异显著(p <0.05)。下同。
Different small letters indicate Duncan significant difference at p <0.05.The same below.

2.2 不同矮化中间砧对华红苹果总酚含量的影响

在果实发育前期,不同矮化中间砧华红果皮总酚的含量存在显著差异,随着果实成熟,差异逐渐减小,在成熟期,差异并不显著(图2)。6 月9 日,SH3组合、SH38 组合和MD001 组合果皮总酚含量低于对照,其中MD001 组合在p <0.05 达到显著水平,为5 254.81 mg·kg-1;CG24 组合、Mac9 组合和CX5组合高于对照,其中CG24 组合显著高于对照,为7 224.33 mg·kg-1。6 月29 日,SH3 组合、SH38 组合和MD001 组合低于对照,差异不显著;CG24 组合、CX5组合和Mac9组合高于对照,差异不显著。7月20日,不同矮化中间砧组合果皮总酚含量均低于对照;除Mac9 组合外,其他组合均显著低于对照。8月8 日,CG24 组合显著高于对照,其他组合差异不显著。随着果实发育,不同矮化中间砧组合果皮总酚含量与对照无显著性差异。在果实成熟期,果皮总酚含量 SH38>CG24>SH3>CK>MD001>CX5>Mac9;SH38 组合最高,为1486.44 mg·kg-1,Mac9 组合最低,为1 117.50 mg·kg-1,两者之间达到显著差异水平,其他组合之间差异不显著。

图2 不同矮化中间砧对华红果皮总酚含量的影响
Fig.2 Effect of different interstocks on content of total phenolics in peel of Huahong apple

不同矮化中间砧在果实发育的前期对果肉总酚的含量有较为明显的影响,随着果实的成熟,果肉总酚含量差异逐渐减小,在果实成熟期无显著差异(图3)。在6月9日,6个中间矮化砧组合果肉总酚含量均低于对照,其中,SH3 组合、SH38 组合和MD001组合显著低于对照,MD001 组合最低,为3 579.34 mg·kg-1,对照最高,为4 466.904 mg·kg-1。6 月29日,CG24 组合和Mac9 组合果肉总酚含量略高于对照组,差异不显著。其余四个组合均低于对照,其中,SH3 组合达到显著水平。在随后的果实发育过程中,除了9 月18 日SH38 组合显著低于对照外,其他四个时期矮化中间砧组合和对照果肉总酚含量的差异逐渐缩小,差异均不显著。在成熟期,果肉总酚含量MD001>SH38>CK>CX5>CG24>Mac9>SH3,最高为358.24 mg·kg-1,最低为308.86 mg·kg-1

图3 不同矮化中间砧对华红果肉总酚含量的影响
Fig.3 Effect of different interstocks on contents of total phenolics in flesh of Huahong apple

2.3 不同矮化中间砧对华红苹果多酚组分动态含量的影响

对不同矮化中间砧组合华红果实发育期果皮和果肉多酚组分检测(图4),共检测出21 种组分,果皮、果肉共有组分为8种,分别为原花青素B2、原花青素C1、绿原酸、根皮苷、表儿茶素、3-羟根皮素2'-木糖葡萄糖苷、根皮素2'-木糖苷和槲皮素3-鼠李糖苷;槲皮素3-芦丁糖苷、槲皮素3-半乳糖苷、槲皮素3-葡萄糖苷、槲皮素3-木糖苷、槲皮素3-未知1、槲皮素3-阿拉伯糖苷、槲皮素3-未知2、矢车菊素3-阿拉伯糖苷和矢车菊素3-半乳糖苷共9种组分仅在果皮中检测出;3-羟根皮素2'-葡萄糖苷、5-O-p-香豆酰奎宁酸、隐绿原酸和4-O-p-香豆酰奎宁酸共4 种组分仅在果肉中检测出。其中果皮中矢车菊素3-阿拉伯糖苷和矢车菊素3-阿拉伯糖苷仅于10 月8 日检测出。

图4 不同矮化中间砧华红苹果多酚组分在果皮和果肉中的动态变化趋势
Fig.4 Dynamic change trend of polyphenol components in peel and flesh of Huahong apple with different dwarf interstocks

左上方数字代表趋势类型,左下方数字代表在该趋势下的指标数量。
The upper left figure represents the trend type,and the lower left figure represents the number of indicators under the trend.

利用6种矮化中间砧组合和对照华红皮中15个多酚组分、肉中12个多酚组分、7个时期进行动态趋势分析,共计289 个指标,设定20 个趋势,通过分析得出8个趋势,趋势0和趋势1占主导地位。多酚组分的变化趋势主要为随着果实成熟逐渐降低(趋势0)和在果实发育阶段先降低,随着果实成熟增高(趋势1),不同矮化中间砧组合对果皮、果肉多酚组分的动态变化有影响。

果皮、果肉的槲皮素3-鼠李糖苷、表儿茶素和原花青素B2,果皮槲皮素3-木糖苷、槲皮素3-阿拉伯糖苷、槲皮素3-未知2、3-羟根皮素2'-木糖葡萄糖苷、根皮素2'-木糖苷和根皮苷,果肉隐绿原酸、4-Op-香豆酰奎宁酸、3-羟根皮素2'-葡萄糖苷、绿原酸含量随着果实发育逐渐降低(趋势0);果皮槲皮素3-半乳糖苷含量,果肉根皮苷含量在果实发育阶段先降低,随着果实成熟升高(趋势1);果肉原花青素C1 含量在果实发育过程中先下降后上升,在果实发育中期达到峰值后继续下降,果实成熟期含量升高(趋势5)。

不同矮化中间砧对砧木组分的变化趋势有一定程度的影响。SH38组合、MD001组合、CX5组合果皮槲皮素3-葡萄糖苷含量,CG24 组合、SH3 组合、MD001 组合和CX5 组合果皮绿原酸含量,SH38 组合和CK组合果肉5-O-p-香豆酰奎宁酸,CG24组合、SH38组合、SH3组合、Mac9组合、CX5组合和CK组合果皮原花青素B2含量,果肉3-羟根皮素2'-木糖葡萄糖苷和根皮素2'-木糖苷含量随着果实发育逐渐降低(趋势0);SH38 组合、SH3 组合、MD001 组合、Mac9 组合、CX5 组合和CK 组合果皮槲皮素3-芦丁糖苷含量,SH3 组合果皮槲皮素3-葡萄糖苷含量,SH38组合、SH3组合、CX5组合和CK组合果皮槲皮素3-未知1 含量,SH38 组合、Mac9 组合和CK 组合果皮绿原酸含量,SH3组合、MD001组合和Mac9组合果肉5-O-p-香豆酰奎宁酸含量,随着果实发育前期先降低,随着果实成熟增高(趋势1);CG24 组合果皮槲皮素3-芦丁糖苷含量随着果实发育先升高,在果实发育中期逐渐降低,随着果实成熟迅速升高(趋势16),果皮槲皮素3-葡萄糖苷含量随着果实发育先降低,后升高,在果实发育中期趋于平稳,随着果实发育逐渐降低,在果实发育成熟时升高(趋势8);对照组合果皮槲皮素3-葡萄糖苷含量在果实发育前期先升高后降低,在果实发育中期之后都趋于稳定(趋势13);Mac9组合果皮槲皮素3-葡萄糖苷含量,CG24 组合、MD001 组合和Mac9 组合果皮槲皮素3-未知1 在含量果实发育前期较为稳定,随着果实发育成“W”状变化趋势(趋势10);CG24 和CX5组合果肉5-O-p-香豆酰奎宁酸含量,MD001组合果肉3-羟根皮素2'-木糖葡萄糖苷含量和根皮素2'-木糖苷含量随着果实发育迅速下降,在果实发育期缓慢上升后稳定,在果实成熟期逐渐降低(趋势2)。

2.4 不同矮化中间砧对华红苹果成熟期多酚组分含量的影响

2.4.1 不同矮化中间砧对华红苹果成熟期果皮多酚组分含量的影响 对华红成熟期(10月8日)果皮多酚组分分析(表2),不同矮化中间砧对华红果皮槲皮素3-半乳糖苷含量影响最小,相关变异系数为0.48%,对矢车菊素3-阿拉伯糖苷含量影响最大,相关变异系数为86.25%。

表2 不同矮化中间砧对华红苹果果皮多酚成分含量的影响
Table 2 Effects of different dwarfing interstocks on the content of polyphenols in the peel of Huahong apple
w/(mg·kg-1

注:不同小写字母表示经Duncan 检验差异显著(p <0.05),下同。
Note:Different small letters indicate Duncan significant difference at p <0.05.The same below.

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在果皮黄酮醇中,SH3 组合中槲皮素3-芦丁糖苷、槲皮素3-葡萄糖苷和槲皮素3-未知1 含量显著高于对照,分别较对照提高103.39%、51.84%和107.90%;MD001组合和Mac9组合槲皮素3-木糖苷和槲皮素3-未知2含量显著低于对照;CG24组合槲皮素3-未知2、槲皮素3-木糖苷、槲皮素3-阿拉伯糖苷和槲皮素3-未知2含量显著低于对照,SH38组合槲皮素3-未知2含量显著低于对照。在羟基肉桂酸中,SH38 组合绿原酸含量较对照提升12.21%。在黄烷醇中,就表儿茶素含量而言,SH38 组合和MD001 组合较对照提高12.2%和3.35%,无显著性差异。就原花青素B2和原花青素C1而言,CG24组合和SH38 组合含量显著低于对照,MD001 组合含量分别较对照提升11.39%和2.07%。在二氢查耳酮中,CG24 组合的3-羟根皮素2’-木糖葡萄糖苷和根皮素2’-木糖苷含量最高,分别较对照提高11.39%和1.60%;MD001组合根皮苷含量最高,较对照提升28.54%。在花青苷中,不同矮化中间砧均提高果皮矢车菊素3-阿拉伯糖苷和矢车菊素3-半乳糖苷的含量,其中SH38 组合显著高于对照,分别较对照提升578.7%和67.9%,说明SH38组合可以显著促进华红的着色。

2.4.2 不同矮化中间砧对华红苹果成熟期果肉多酚组分含量的影响 对华红成熟期(10月8日)果肉多酚组分分析(表3),不同矮化中间砧对果肉4-O-p-香豆酰奎宁酸含量影响最大,相关变异系数为114.24%,对原花青素B2和原花青素C1含量影响最小,相关变异系数分别为2.41%和2.35%。

表3 不同矮化中间砧对华红苹果果肉多酚成分含量的影响
Table 3 Effects of different dwarfing interstocks on contents of polyphenol in flesh of Huahong apple w(/mg·kg-1

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在黄烷醇中,MD001 组合和SH38 组合果肉表儿茶素含量分别较对照提升22.40%和11.90%,CG24 组合、SH3 组合、SH38 组合和Mac9 组合原花青素B2 含量显著低于对照。在黄酮醇中,CG24 组合、SH3 组合、MD001 组合、Mac9 组合和CX5 组合槲皮素3-鼠李糖苷含量分别较对照提升29.57%、25.42%、22.51%、6.30%和34.53%。在羟基肉桂酸中,MD001 组合绿原酸、隐绿原酸和4-O-p-香豆酰奎宁酸含量分别较对照提升41.27%、11.17%和112.50%;Mac9 组合隐绿原酸和4-O-p-香豆酰奎宁酸含量分别较对照提高19.15%和26.04%;CG24 组合和CX5 组合隐绿原酸含量分别较对照提升21.28%和3.19%;SH3 组合4-O-p-香豆酰奎宁酸含量较对照提高44.79%。在二氢查耳酮中,MD001组合根皮素2'-木糖苷、3-羟根皮素2'-木糖葡萄糖苷、3-羟根皮素2'-葡萄糖苷和根皮苷含量分别较对照提高18.25%、19.94%、12.57%和73.44%;SH38 组合3-羟根皮素2'-木糖葡萄糖苷、根皮素2'-木糖苷和根皮苷含量分别较对照提高8.72%、21.90%和55.73%;SH3 组合根皮素2'-木糖苷和根皮苷含量较对照提高6.57%和102.08%;CG24 组合、Mac9 组合和CX5组合根皮苷含量分别较对照提高2.60%、177.08%和31.77%。

3 讨 论

3.1 华红果实发育过程中总酚及组分含量的动态变化

类黄酮是多酚物质的一类,前人研究表明苹果中含有100多种类黄酮化合物[22-24]。研究表明长富2号苹果不同果实部位之间类黄酮物质的含量和组分存在明显差异[25]。在本试验中,果皮检测出17种类黄酮组分、果肉则检测出12 种。其中8 种类黄酮物质果皮和果肉共有,9 种多酚组分只在果皮中检测出,4种只在果肉中检测出,说明苹果果实不同部位类黄酮组分含量不同。有两种槲皮素组分未探明其具体名称,有待采用更先进的手段进行鉴定。

苹果生长发育前期,总酚含量最高,随时间的变化,总酚含量不断降低[26]。本研究中,华红随着果实的发育,果皮和果肉总酚含量逐渐降低,发育早期,总酚含量迅速下降,果实成熟期趋于稳定或略有下降,果皮中总酚含量显著高于果肉,总酚含量在果实不同部位的含量变化趋势总体呈现一致性。随着华红果实的成熟膨大,大部分类黄酮物质在果皮和果肉中减少,是总酚在成熟期呈现下降趋势的主要原因。果皮和果肉槲皮素3-鼠李糖苷、表儿茶素和原花青素B2含量在果实发育初期最高,果实成熟时含量最低,果肉原花青素C1含量随着果实的成熟膨大而变化,在果实发育中期先下降后上升,随后在果实发育后期下降,在成熟期有所上升,本研究中的华红多酚组分变化趋势和津轻基本相同[27],苹果根皮苷含量随着果实成熟逐渐降低[28]。本研究中,华红果肉的根皮苷含量在果实成熟升高,但果皮根皮苷含量则随着果实成熟而降低。

3.2 不同矮化中间砧木对华红总酚及组分和含量的影响

矮化中间砧对苹果酚类物质的影响是多方面的,解贝贝等[29]对嘎拉苹果的研究发现,嫁接到不同砧木上会对酚类物质含量产生影响,其中将嘎拉嫁接到A1 上总酚含量最高。矮化中间砧可以整体上提高某些苹果品种的果实功能性成分含量(总糖、可溶性固形物和酚类物质),Mac9 和CG24 嫁接的蜜脆果实功能性成分含量的综合评价最高[30]。在本研究中,CG24 组合和Mac9 组合在果实发育早期的总酚含量高于其他矮化中间砧组合,但随着果实的动态发育,果实成熟期,CG24 组合和Mac9 组合的果肉总酚含量低于对照和其他组合,但CG24 组合果皮总酚含量却高于对照和Mac9 组合,CG24 组合黄烷醇和黄酮醇组分含量随着果实的发育,下降幅度明显高于其他组合和对照,是其总酚含量下降的主要原因。另外,CX5组合果实成熟期果皮、果肉总酚含量均低于对照,综合来看,推测Mac9 组合和CX5组合较低的总酚水平,可以提高华红抗褐化能力。Kviklys等[31]研究认为砧木对苹果槲皮素类、表儿茶素和绿原酸含量的影响较小,在本试验中,不同矮化中间砧的华红果皮、果肉表儿茶素和绿原酸含量无显著差异,槲皮素类组分的含量有不同程度提高。同时显著提高果皮花青苷的含量。王大江等[30]报道不同矮化中间砧蜜脆花青苷和黄酮醇组分的含量均有不同程度的提高,宋伊真等[13]研究发现同一发育时期,同一苹果品种,因砧木不同其果皮和果肉的总酚和类黄酮含量存在差异,结合本试验,推测砧木可以对苹果果实酚类物质的含量产生一定的影响。可进一步测定不同矮化中间砧和品种组合对苹果多酚组分及含量的影响,阐明引起嫁接苹果酚类物质组分及含量差异的原因。

果实的颜色主要由花青苷含量调控,花青苷含量是随着果实的发育逐渐增加[32],仅存在于红色苹果果皮中[33]。苹果中已报道了花青苷类物质有矢车菊素3-半乳糖苷、矢车菊素3-阿拉伯糖苷和矢车菊素3-木糖苷共计3 种花青苷[34]。华红主要检测出矢车菊素3-半乳糖苷、矢车菊素3-阿拉伯糖苷。Fallahi等[35]发现嫁接矮化中间砧的Scarlet Gala苹果颜色有显著变化,其中嫁接M9 和M26 中间砧果实较红。砧木影响果实转色期花色素苷合成途径中苯丙氨酸氨解酶(PAL)、查尔酮异构酶(CHI)、类黄酮葡糖苷转移酶(UFGT)3个酶活性,从而影响花青苷合成,引起果皮色泽表现[36]。本研究发现不同矮化中间砧的华红成熟期果实花青苷含量均有提升,以Mac9 组合和SH38 组合最为显著,两个组合提升了果皮矢车菊素3-阿拉伯糖苷和矢车菊素3-半乳糖苷含量,进而提升果皮花青苷含量。推测矮化中间砧可以促进华红着色,提升着色度以Mac9 组合和SH38组合最优,可以进一步对嫁接不同矮化中间砧华红着色期的相关基因进行定量表达分析,解释矮化中间砧引起的苹果着色机制。

3.3 多酚在华红苹果果肉抗褐化中的可能作用

多酚作为苹果的次级代谢产物,是一类结构多样且活性各异的化合物集合,是苹果中的主要活性成分,具有抗逆、抗氧化等重要的生理功能[27]。苹果多酚是褐变反应的底物之一,苹果酶促褐变是多酚氧化酶(PPO)引起的,酚类物质被氧化后经非酶促聚合,生成黑色素,果肉褐化,苹果总酚含量与苹果褐化程度呈正相关[37]。不同品种的苹果对褐变的敏感程度不同,富士系品种对褐化较为敏感,金冠、红星和青香蕉等品种贮藏时不易发生果肉褐变[38]。乜兰春等[39]研究发现相同贮藏条件下,富士贮藏中褐变度最高,其次为新红星,金冠最低。不同苹果果肉总酚含量如下:富士726.0 mg·kg-1[4]、乔纳金1100 mg·kg-1[40]、旭428.1 mg·kg-1、科特兰536.5 mg·kg-1、红星445.8 mg·kg-1和金冠370.3 mg·kg-1[41],本研究中,华红成熟期果肉总酚含量为308.86~358.24 mg·kg-1,低于容易褐化鲜食品种富士,与抗褐化品种金冠接近。

在多酚组分中,绿原酸和儿茶素被认为是引起褐变的主要底物[42]。果实的褐变度与表儿茶素、儿茶素及绿原酸含量呈显著正相关,相关性最高的是表儿茶素,其次为绿原酸,表儿茶素含量较低苹果抗褐化能力则越强[43]。富士果肉绿原酸和表儿茶素含量分别为550 mg·kg-1和190 mg·kg-1[4],金冠为153.6 mg·kg-1和65.8 mg·kg-1[41]。成熟期华红果肉中表儿茶素和绿原酸含量分别为28.18~44.72 mg·kg-1和24.64~46.13 mg·kg-1,表儿茶素和绿原酸含量明显低于褐化品种,推测华红苹果果肉抗褐化的能力可能来自低的多酚含量水平,表现在较低的表儿茶素和绿原酸含量水平。

4 结 论

在华红果实中,总酚含量随着果实的发育逐渐降低,果皮、果肉在7个发育时期总酚含量均存在差异,果皮中总酚含量明显高于果肉;在果实成熟期,不同矮化中间砧组合的多酚组分相似,都以原花青素、表儿茶素和绿原酸为主;大多数多酚组分的含量随着果实成熟逐渐降低,少部分则在果实发育阶段先降低,随着果实成熟增高,不同矮化中间砧组合对果皮、果肉多酚组分的动态变化有影响。果实成熟期,Mac9 组合和SH38 组合显著提高华红花青苷含量,进而促进果实着色;SH38组合和MD001组合对果皮、果肉多酚组分的含量提升较多。综合来看,SH38 组合可考虑作为提高华红苹果功能性栽培的首选中间砧。

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Effects of different dwarfing interstocks on the total phenols and polyphenol components in Huahong apple

LI Qingshan1, 2, GAO Yuan2, WANG Kun2, SUN Simiao2, LU Xiang1,2, LI lianwen2, FENG Jianrong1*,WANG Dajiang2*

(1College of Agriculture,Shihezi University/The Key Laboratory of Special Fruits and Vegetables Cultivation Physiology and Germplasm Resources Utilization, Xinjiang Production and Construction Corps, Shihezi 832000, Xinjiang, China;2Research Institute of Pomology,Chinese Academy of Agricultural Sciences/Key Laboratory of Horticultural Crops Germplasm Resources Utilization,Ministry of Agriculture and Rural Affairs,Xingcheng 125100,Liaoning,China)

Abstract:【Objective】The dynamic effects of six different dwarfing interstocks on the total phenols and polyphenol components in Huahong apple were studied in order to provide a basis for selection and application of apple dwarf interstocks and fruit production with high quality.【Methods】The rootsock was Malus baccata (L.) Borkh., and the interstocks were CG24, SH38, SH3, MD001, Mac9 and CX5.The scion variety Huahong was grafted on the interstocks,and the trees of Huahong grafted on M.baccata (L.) Borkh. were used as control (CK). The components and contents of polyphenols in the peel and flesh of Huahong fruits were analyzed at seven periods during Huahong apple growth using High Efficiency Liquid Chromatography and Mass Spectrometry, and the content of total phenols was determined by Folin-Ciocalteu method.【Results】The content of total phenols in the peel of Huahong fruits was significantly higher than that in the flesh, the content of total phenols in the peel and flesh decreased gradually during the period of the fruit development. However, the periods of the lowest content of total phenols were different in the peel and flesh, it was the 8th October in the flesh and the 29th August in the peel. The content of polyphenols in the peel and flesh decreased gradually with fruit development and maturity or decreased first at the stage of fruit development, and then increased at the maturity stage.Different interstocks had different effects on the dynamic changes of polyphenols in the peel and flesh.At maturity stage,the content of total phenols in the peel of Huahong fruits from high to low was SH38 >CG24 >SH3 >CK >MD001 >CX5 >MAC9,and the content of total phenols in the flesh was MD001 >SH38 >CK >CX5 >CG24 >MAC9 >SH3. During the period of fruit ripening,SH3,SH38 and Mac9 significantly increased the content of flavonols in the peel,MD001 significantly increased the content of hydroxycinnamic acids in the flesh. The content of quercetin-3-alactoside was the highest in the peel and the content of procyanidin B2 was the highest in the flesh at the stage of fruit maturation.At the stage of fruit maturation,the contents of epicatechin and chlorogenic acid of Mac9 and CX5 were low relatively compared with the others in the flesh;the anthocyanins of SH38 and Mac9 were high relatively compared with the others in the peel.The polyphenol components mainly decreased gradually with fruit maturity.With the increase of fruit maturity,different dwarf intermediate rootstocks had an impact on the dynamic changes of polyphenol components in the peel and flesh.The content of Quercetin 3-rhamnoside,Epicatechin and Procyanidin B2 in the peel and flesh and the content of Quercetin 3-xyloside,Quercetin 3-arabinoside,Quercetin 3-unknown 2,3-Hydroxyphloem 2’-xylose glucoside, Phloem 2’-xyloside and Phloridzin in the peel, the content of 4-Dicaffeoylquinic acid, 4-O-p-coumaroyl quinic acid, 3-Hydroxyphloretin-2-glucoside and Chlorogenic acid in the flesh rapidly declined during fruit development,and then it kept stable basically.But the contents of Quercetin3-alactoside in the peel and Phloridzin in the flesh were accumulated as fruit ripening.The content of procyanidin C1 in the flesh decreased first and then increased during the period of fruit development,continued to decrease after reaching the peak in the middle of fruit development,and increased at fruit maturity. Different dwarfing interstocks had some effects on the change trend of polyphenol components. The content of Quercetin3-glucoside in the peel of the fruits on SH38, MD001 and CX5,the content of Chlorogenicacid in the peel on CG24,SH3,MD001 and CX5,the content of 5-O-p-coumaroyl quinic acid in the peel on SH38 and CK, the content of 3-Hydroxyphloretin-2-xylglucoside,Phloretin-2-xyloside in the flesh and the content of Procyanidin B2 in the peel on SH38, SH3, Mac9,CX5 and CK gradually decreased with fruit development. But the content of Quercetin3-rutinoside in the peel on SH38, SH3, MD001, Mac9, CX5 and CK, the content of Quercetin3-glucoside in the peel on SH3, the content of Quercetin3-unknown1 in the peel on SH38, SH3, CX5 and CK, the content of Chlorogenic acid in the peel on SH38,Mac9 and CK,the content of 5-O-p-coumaroyl quinic acid in the flesh on SH3, MD001 and Mac9 first decreased at the early stage of fruit development and increased with fruit maturity.【Conclusion】The content of total phenols of Huahong fruits decreased gradually during the development of fruits, and the content of total phenols in the peel was significantly higher than that in the flesh. The content of polyphenolic components of the peel and flesh decreased rapidly during the beginning of fruit development, then it kept stabile basically, but decreased gradually during fruit ripening, however, the content of quercetin3-rutinoside, quercetin3-alactoside, quercetin3-unknown 1 and quercetin3-glucoside in the peel,the content of phloridzin and procyanidin C1 in the flesh were accumulated with fruit ripening.The combination of Mac9 and SH38 significantly increased fruit coloration,SH38andMD001couldincresethepolyphenolcontentinthepeelandflesh.Inconclusion,SH38 couldbeconsideredassuitableinterstockforcultivationofHuahongappletoproducefunctionalfruits.

Key words:Apple;Huahong;Dwarfing interstocks;Phenols;Polyphenolic components

中图分类号:S661.1

文献标志码:A

文章编号:1009-9980(2022)07-1191-12

DOI:10.13925/j.cnki.gsxb.20210612

收稿日期2021-11-29

接受日期:2022-03-04

基金项目中国农业科学院科技创新工程项目(CAAS-ASTIP-2018-RIP-02);财政部和农业农村部:国家现代农业产业技术体系资助项目(CARS-27)

作者简介李青山,男,硕士,主要从事果树种质资源研究。Tel:15623906211,E-mail:sichenff@163.com

*通信作者Author for correspondence.Tel:13031329662,E-mail:fengjr102@sohu.com;Tel:13842953961,E-mail:dajiang0101@126.com