番茄(Lycopersicon esculentum),又称西红柿,成熟的果实呈球形,颜色艳丽,酸甜可口,有特殊风味。经常食用番茄有益于人体健康[1],特别是番茄红素极具营养价值,具有增强免疫力[2]、保护神经[3]、控制血糖[4]等功效。除了番茄果肉有较高的经济价值外[5],它的副产物也有极大的开发潜力,可有效预防胆固醇过高和调节肠道菌群[6],也具有一定的抗菌能力[7]。但是由于含水量高[8]、果实的呼吸作用[9]等,番茄会面临采摘后易腐烂变质的问题,极大地降低了番茄的价值。为避免因番茄变质造成大量浪费,需要对如何延长番茄的贮藏期、保持贮藏期间番茄的品质进行研究。
目前,果蔬的贮藏保鲜方式很多,并且在不断改进完善。采摘后的番茄常通过低温贮存来延长番茄保质期,在一定程度上能有效增强番茄的贮藏性[10-11],也可以使用保鲜剂进行保鲜,例如1-甲基环丙烯频繁地被使用在果蔬的贮藏中[12-13],通过有效地抑制乙烯作用达到保鲜目的[14]。除此之外,臭氧保鲜和气调保鲜也在番茄贮藏中有一定的应用,但仍存在一些局限性,如气调保鲜的气体比例难以确定[15]、保鲜剂用量难以把控[16]等,使用不当都会致使果蔬无法延长保质期,甚至加速腐烂。
植物提取物大多是从天然植物中提取纯化得到的具有生物活性的物质,例如挥发油、多酚、黄酮及生物碱[17]等,因其安全无毒且抗菌防腐被广泛研究[18],可在一定程度上代替农药、保鲜剂[19]等。使用天然植物提取物进行果蔬的贮藏保鲜,安全无毒、低残留,可有效避免化学保鲜剂高毒、高残留等问题。本文探究了影响番茄贮藏的因素和植物提取物保鲜技术机理,以及其在番茄贮藏保鲜的应用,并且为植物提取物保鲜技术今后的发展提供依据。
果蔬在贮藏期间会发生一系列品质变化[20],包括颜色变化、表面皱缩、内部成分减少或增加,并且会产生腐烂变质,这种现象大多是由微生物、酶、自身物理化学反应等造成[21]。除微生物的影响外,温度、品种、成熟度以及运输、包装方式都与果蔬品质的维持息息相关,若忽视这些因素的影响,会增加番茄的腐烂程度,减短番茄的保存时间。
番茄成熟后皮薄且柔软多汁,容易形成机械损伤,创伤表面易受到病原菌的感染[22],在贮藏期间,灰霉病、黑霉病、果腐病等是困扰番茄贮藏的病害,会造成番茄产量的巨大损失[23]。
灰霉病由灰葡萄孢(Botrytis cinerea)引起,易感染侵害含水量高的果蔬[24],传染力强、传播速度快,被侵染的番茄果实变软变色,最终表面还会附着灰褐色霉层。彭家丽等[25]给番茄植株接种灰霉病菌,发现会导致番茄叶片中酶和淀粉含量的变化,并伴随氧化损伤等现象。除此之外,番茄感染灰霉病后会大量产生乙烯,加速果实的衰老和腐烂[26]。
病原菌链格孢(Alternaria alternata)经常会引起番茄黑霉病,一般危害番茄的茎和果实,果实最后会产生灰黑色霉层。除了致腐作用,链格孢霉菌在一定条件下会产生毒素[27],危害人体健康。
果腐病的病原菌会使番茄颜色形态发生变化[28]。最常见的青霉果腐病由扩展青霉菌(Penicillium expansum)引发,会使番茄出现绒状暗绿色、近球形或椭圆形的菌落。乳酸镰刀菌(Fusarium lactis)从腐烂番茄中分离,具有致病性,可使番茄果实加速腐烂[29]。还有引发根腐病的尖孢镰刀菌(Fusarium oxysporum),同样易导致番茄果实塌陷软化[30]。
温度对番茄贮藏至关重要,会影响果皮的弹性、酶活性、果胶、细胞内水分等,长时间高温甚至会造成番茄果皮破损裂开[31],会加剧微生物的侵染。而且贮藏温度不会保持一成不变,温度的起伏也会影响果蔬的贮藏情况,刘佳等[32]证明了温度波动最小时能保持番茄的良好品质,可延长保质期2 d~4 d。低温可有效减缓果蔬衰老,李金娜等[33]研究发现5℃是番茄最佳保存温度,此温度条件下番茄能保持良好形态及颜色,且不易软化,较大程度避免营养的流失,在此温度下可再延长贮藏期5 d~7 d。曲劲尧等[34]比较了接种链格孢霉分生孢子的番茄15 d内在不同温度贮藏的情况,表明低温更容易阻碍链格孢霉的生长。但不适宜的低温同样不利于番茄的贮藏,番茄灰霉病最易在低温、高湿的环境中侵染[35]。
番茄贮藏期的品质及生物活性的保持与番茄的品种及采摘时的成熟度密切相关[36]。毛方华等[37]通过比较智能温室培养的番茄贮藏性,发现农科2号果实在贮藏期间表现出更好的品质,农科1号的腐烂率更低,有更长的保质期。王倩等[38]探究了不同品种番茄贮藏的差异,低温[(10.0±0.2)℃]环境中贮藏,“欧盾”品种番茄在贮藏期内能保持良好硬度,且乙烯释放量较小,能有效降低相关酶活性。Olveira-Bouzas等[39]比较分析不同成熟度对番茄贮藏的影响,将收获的番茄按颜色分为成熟度不同的两组,在相同的条件下贮藏,测定其指标,结果表明与成熟的番茄相比,未成熟的番茄有更长的贮存期。
选择适当的运输与包装形式,能尽量避免磕碰,减少番茄机械损伤,消除番茄表面破损有利于番茄保存,并且能减少微生物入侵的几率[40]。高雅等[41]探究不同运输和包装方式对贮藏28 d后番茄的影响,结果显示,厢货的运输方式更好,可以减缓番茄的不良变化,而对于包装方式,选择微孔保鲜膜处理番茄再用瓦楞纸箱包装的效果更好,更有利于番茄贮藏。
导致果蔬腐烂的重要因素为微生物的侵染,而植物提取物具有抗菌性[42-43],可有效防止微生物造成的腐烂变质[44-46],具体应用如表1所示。
表1 植物提取物对微生物的抑制效果
Table 1 Inhibitory effect of plant extracts on microorganisms
注:-表示所引文献中未提及该部分内容。
序号 种类 抑菌主要成分 抑制微生物 参考文献1 香漆柏精油 α-蒎烯、β-石竹烯、环氧石竹素 灰霉病菌 [42]2 牛蒡和苔干复配提取物 - 大肠杆菌、金黄色葡萄球菌、蜡样芽孢杆菌 [43]3 花椒提取液 - 扩展青霉、灰葡萄孢、链格孢菌 [44]4肉桂精油 反式肉桂醛、丁香酚、芳樟醇 粉红单端孢菌、灰葡萄孢菌、链格孢菌、扩展青霉菌 [45]-大肠杆菌、金黄色葡萄球菌 [47]-链格孢菌 [48]-软腐病菌 [49]5 辣椒叶片提取物 没食子酸、绿原酸、儿茶素、咖啡酸、对香豆酸、阿魏酸链格孢菌 [50]6木醋酸 - 灰霉病原菌、番茄早疫病原菌、番茄黑斑病原菌、番茄枯萎病原菌[51]7 翅果竹芋精油 石竹烯 曲霉、尖孢镰刀菌、白地霉 [52]8山香精油 愈创木酚9 闭果桐精油 β-香豆素10 丁香精油 丁子香酚 黑曲霉、尖孢镰刀菌 [53]丁香酚和石竹烯 灰葡萄孢霉、粉红单端孢霉、串珠镰刀菌 [54]11 当归茎叶精油 2,4,5-三甲基苯甲醛、α-蒎烯 粉红单端孢菌、硫色镰刀菌、互隔交链孢菌、扩展青霉菌 [55]12 香叶树提取物 金丝桃苷、芦丁、异槲皮苷、扁蓄苷、野黄芩苷、儿茶素、表儿茶素灰霉病菌 [56]13 橙皮提取物 - 毛霉、曲霉、叶霉、番茄灰霉 [57]-黑曲霉、桔青霉 [58]14 芍药花提取物 - 革兰氏阳性菌、革兰氏阴性菌、酵母、枯草芽孢杆菌 [59]15 薰衣草精油 - 金黄色葡萄球菌、大肠杆菌、单核细胞增生李斯特菌 [60]16 芸香精油 2-十一烷酮、2-壬酮 霉菌、酵母、嗜氧菌嗜温细菌 [61]17 百里香精油 - 大肠杆菌 [62]百里香酚 - 灰霉病菌 [63]18 连翘精油 - 大肠杆菌、金黄色葡萄杆菌 [64]
Pane等[50]使用野生辣椒提取物,发现提取物中富含酚类物质,能够有效抵抗链格孢菌对番茄的感染。杨苏苏等[51]验证了22种植物提取物对番茄常见病害的防治作用,其中木醋酸对番茄灰霉病原菌、早疫病原菌、黑斑病原菌、枯萎病原菌的抑制作用可达75%以上。Dania等[52]使用水蒸气蒸馏法提取的翅果竹芋、山香、闭果桐精油,抑制了3种番茄果实腐烂病菌(曲霉、尖孢镰刀菌和白地霉)的生长,减少番茄的腐烂。丁香精油中的丁子香酚也能用于去除番茄病害[53]。陈梅[54]针对会使番茄引起灰霉病的灰葡萄孢霉,选取丁香精油作为研究对象,对番茄采后病原菌进行精油熏蒸,结果表明丁香精油可以在一定程度上降低灰霉病的危害。当归茎叶精油也能有效降低粉红单端孢的生长繁殖[55]。
由于植物提取物中含有多种化合物[65],所以它对果蔬的保鲜机理较为复杂,作用机制尚不明确,需要对其进行更深入探究。就目前来看,植物提取物的抑菌作用主要是通过影响细胞结构或细胞内活动来限制微生物的生长[66]。牡丹花蕾提取物的抑菌防腐机理可以主要归结于提取物对细菌细胞膜成分及其通透性、流动性、完整性的影响,以及破坏DNA的合成,影响相关基因的表达,同时抑制相关酶活性和相关毒力因子的转录,进而导致细菌死亡[67]。鼠尾草精油可影响细胞壁、细胞膜的形态,造成破裂、下陷、扭曲等,并且会减少三磷酸腺苷(adenosine triphosphate,ATP)和胞内核酸含量,进而达到抑菌效果[68]。山苍子精油破坏细胞膜结构,使胞内大分子物质泄漏,影响细胞呼吸作用,抑制三羧酸循环(tricarboxylic acid cycle,TCA)和降低α-酮戊二酸脱氢酶等相关酶的活性,降低ATP含量并抑制相关酶活性,同时破坏DNA结构,影响其基因表达[69]。依据各类植物提取物的研究可总结归纳其保鲜机理如图1所示。
图1 植物提取物的保鲜抑菌机理
Fig.1 The fresh-keeping and antibacterial mechanism of plant extracts
植物提取物保鲜技术相较其他保鲜技术,安全、无毒、无残留、成本较低、实施更加方便。
植物提取物大多通过超临界CO2萃取法[70]、溶剂萃取法[71]、微波辅助提取法[72]等从各种植物中提取纯化,可用于果蔬的贮藏,在番茄贮藏研究中具有很高的价值,植物提取物保鲜技术在番茄贮藏中的应用见表2。
表2 植物提取物保鲜技术在番茄贮藏中的应用
Table 2 Application of plant extract preservation technology in tomato storage
序号 提取物名称 使用条件 使用效果 参考文献1 山苍籽精油 150 μL/L的山苍籽精油,熏蒸24 h,(25±1)℃贮藏 与对照组相比,以好果率为标准,可延长贮藏期8 d [73]2 肉桂精油 以0.5%壳聚糖为外层,海藻酸钠与两亲性淀粉包裹的肉桂精油抗菌膜与聚乙烯膜相比,抗菌膜具有一定的保鲜效果,在2周内保持更高的硬度和更低的失重率,具有优异的抗菌活性和保鲜性能[47]在番茄生长时使用可降解保护罩,精油与聚乳酸(polylactic acid,PLA)、聚己二酸对苯二甲酸丁二醇共聚酯(polybutylene adipate terephthalate,PBAT)两种可降解材料512 μg/mL 伊枯草菌素 A(iturin A)与 1%(体积分数)肉桂精油复配使用与对照组相比,可延缓番茄的发育成熟 [48]与对照组相比,以腐烂率为标准,可延长贮藏期9 d [49]熏蒸法,100 μL/L的肉桂精油 与对照组相比,以好果率为标准,可延长贮藏期10 d [74]3 八角精油 浸泡在八角精油中10 min,后贮藏在25℃中 与对照组相比,以失重率为标准,可延长贮藏期6 d [75]4 枸杞叶多酚 在室温20℃环境下,浸泡于0.4 g/L的枸杞叶多酚溶液与对照组相比,以质量损失率为标准,可延长贮藏期15 d[76]5 橙皮黄酮 1 g/L的橙皮黄酮浸泡保鲜纸,进行番茄包装 与对照组相比,可延长寿命6 d [57]6 柑橘精油 0.8%柑橘精油与1%壳聚糖形成的复合保鲜液处理2 min,于4℃冰箱内贮藏与对照组相比,以腐烂率为标准,可延长贮藏期12 d [77]在25℃下储存,与壳聚糖(2%)的组合作为涂层 有效控制了西红柿的真菌腐烂,可延长贮藏期至8 d [58]7 芍药花提取物 番茄于芍药花提取物(10 mg/mL)浸泡3 min后,于25℃下贮藏与对照组相比,以腐败率为标准,可延长贮藏期6 d [59]
续表2 植物提取物保鲜技术在番茄贮藏中的应用
Continue table 2 Application of plant extract preservation technology in tomato storage
序号 提取物名称 使用条件 使用效果 参考文献8 薰衣草精油 在(25±2)℃下贮藏,使用水包油薰衣草纳米薄膜包裹番茄可延缓番茄成熟,抑制酚类物质氧化,使番茄保持新鲜状态[60]9 芸香精油 在4℃条件下,结合壳聚糖使用 可保持番茄品质,减少腐烂 [61]10 百里香精油 采用超声纳米乳化法制备百里香精油纳米乳 保持番茄品质及硬度 [62]百里香酚 与乙烯清除剂高锰酸钾(KMnO4)结合使用 延缓水分的流失,保持良好的感官品质,延长保质期 [63]11 连翘精油 在25℃、相对湿度65%的条件下,用连翘精油1.50%、壳聚糖2.0%、ε-聚赖氨酸0.50%、Nisin 0.05%制备保鲜液能有效保持番茄品质,延长番茄货架期 [64]连翘提取液 连翘叶提取物与壳聚糖复配物以体积比为5∶1混合浸泡3 min后放置室温(18±2)℃贮藏12 鹰嘴豆壳多糖 鹰嘴豆壳多糖结合羧甲基纤维素制备薄膜,涂于圣女果表面与对照组相比,以失重率为标准,可延长贮藏期6 d [78]可延长番茄保质期,维持其品质 [79]13 桉树精油 桉叶精油微胶囊应用于樱桃番茄26℃贮藏 与对照组相比,可延长贮藏期2 d [80]14 粉红胡椒精油 使用粉红胡椒精油乳液,对番茄进行熏蒸 延缓了樱桃番茄的成熟,维持其良好品质 [81]
在番茄的贮藏保鲜中,较为常见的是肉桂精油、薰衣草精油、月桂精油及丁香精油等。王自鹏等[73]采用熏蒸法处理番茄,隔天测定番茄的果实外部状态、内部变化等相关指标,比较不同组的变化趋势,分析在(25±1)℃下不同剂量的山苍籽精油对番茄的贮藏影响,与其他剂量组相比,150 μL/L山苍籽精油作用最明显,好果率最高,品质最好,可有效延长贮藏期。不同剂量的肉桂精油浸于滤纸上,贴在贮藏盒上对圣女果进行熏蒸,每隔1 d测定圣女果的贮藏指标,100 μL/L的肉桂精油效果最好,可降低果实的失重率和细胞的通透性,保持果实硬度和品质[74]。李姗姗等[75]比较了八角、金银花、香樟叶3种植物的精油对番茄的保鲜贮藏作用,并且设置了蒸馏水对照,八角精油相较其他精油抑制效果最好,可明显降低营养成分的损失,保持相关酶的活性,可延缓番茄的衰老。
除使用精油外,还有其他许多活性成分也可用于贮藏保鲜[56],应用于番茄的贮藏。司宝华等[76]探究枸杞叶多酚对番茄的贮藏保鲜作用,在20℃下,不同剂量的枸杞叶多酚有不同的效果,结果表明0.4 g/L剂量的枸杞叶多酚的保鲜效果最好,可以很好地维持番茄的品质,0.2、0.6 g/L效果次之,0.8 g/L几乎不起作用。王晓君[57]研究了橙皮黄酮的最佳提取工艺,橙皮黄酮提取量为3.96 g/100 g,并探究橙皮黄酮对番茄贮藏期常见微生物的抑制效果和最小抑菌浓度,证明1 g/L橙皮黄酮制备的保鲜纸具有保鲜作用,可有效减弱呼吸强度的影响,推迟呼吸高峰的到来。芍药花提取物[59]也可作为保鲜剂使用在番茄贮藏中,采用乙醇提取法提取芍药花提取物,充分证明提取物具有保鲜抑菌作用,可保持贮藏期内番茄的品质。
较目前来说,植物提取物保鲜也暴露出一些不足。一些植物提取物的易挥发性使它们的保鲜效果大打折扣,并且本身具有味道[82],使用过量时,可能会掩盖食物本身的味道,带来不好的感官体验。这需要加大对提取物使用剂量的把握以及对使用形式进行深入研究。所以对植物提取物保鲜技术的研究不应仅仅局限于提取物本身的探索,还需要对其如何最大限度地发挥作用进行思考。
可食性包装使用安全天然、可降解的材料制备,可代替食品的包装材料,减少对环境的危害[83],并且弥补植物提取物保鲜的缺陷。涂膜是可食性包装最常用的形式之一,壳聚糖最常与植物提取物结合进行涂膜保鲜,具有良好成膜性能、稳定性好,并且在一定程度上能够减少食物水分散失[47]。张莹丽等[77]制备了一种复合涂膜保鲜剂,采用柑橘精油结合壳聚糖,通过对不同剂量的壳聚糖和柑橘精油组合试验,认为0.8%柑橘精油与1%壳聚糖形成的复合保鲜液能显著减缓圣女果的腐败,并维持圣女果的采后品质。Sheikh等[58]同样使用柑橘皮精油结合壳聚糖,结果与之类似。潘婷婷等[78]使用连翘提取液结合壳聚糖,探究对圣女果的保鲜作用,当体积比为5∶1时,保鲜效果最佳,可减缓腐败率和失重率,维持圣女果贮藏期的品质。纳米乳液也常在果蔬贮藏中出现[84],能够增大植物提取物的溶解度,更好地发挥植物提取物的保鲜作用。Sun等[60]探究薰衣草精油对番茄的贮藏保鲜作用,以水包油薰衣草精油纳米乳液的形式,混合明胶制备成保鲜膜,结果表明,制备的保鲜膜可抑制樱桃番茄的变质和微生物的生长,基本可以维持番茄品质。对精油包装进行探索,或许可以减少精油提取成本的损耗,增强精油的保鲜效果。除壳聚糖、纳米乳液材料,还有羧甲基纤维素(carboxymethyl cellulose,CMC)[79]、葡甘露聚糖[85]等材料也经常与植物提取物结合,使番茄具有更长的保质期,减少番茄贮藏时的不良变化。
可食性包装不接触食物表面,也同样可以达到保鲜防腐的效果。Black-Solis等[48]为防止番茄腐烂软化,在番茄生长期间罩上了涂有肉桂精油的可生物降解聚酯网,可以有效防止链格孢菌的侵袭,减少黑霉病的发生,并且可延长番茄保质期,保持番茄的品质。岳淑丽[80]研究桉树精油的保鲜作用,采用包合法用β-环糊精制备桉叶精油微胶囊,可以延缓番茄果实的品质下降,减少维生素C等物质的消耗,可延长番茄贮藏期2 d。Locali-Pereira等[81]研发出一种新型的精油保鲜包装形式,可减少精油的挥发和降低精油本身的味道,将粉红胡椒精油分别包埋在单层(大豆分离蛋白)和双层(大豆分离蛋白+高甲氧基果胶)乳液中,并且使用两种包装方式,一是涂在盒盖表面形成图层,二是装在袋子中贴在盒盖上,同条件下贮藏21 d后,与没有进行精油涂抹的番茄对照,结果表明,精油具有良好的番茄保鲜作用,都可以延缓番茄的成熟,但双层乳液具备更好的稳定性,涂层式的包装方式更加有效。
由于植物来源广,有效成分不稳定,不同提取物的提取工艺不同,这为植物提取物保鲜技术的推广、应用带来巨大阻碍[86]。所以将植物提取物保鲜技术与其他保鲜技术相结合,可有效弥补植物提取物提取难、提取量少的缺陷,甚至事半功倍,能产生更好的保鲜效果。与物理保鲜技术结合是最为常见的保鲜组合,且简便有效。Peralta-Ruiz等[61]联用低温保鲜技术,在4℃的条件下,芸香精油混合壳聚糖应用于番茄贮藏,能有效抑制番茄的不良变化,具有防腐效果。He等[62]使用超声技术辅助百里香精油纳米乳液的保鲜作用,能显著保持番茄的品质,降低微生物污染。蒋梦曦等[49]结合生物保鲜技术,探究肉桂精油的贮藏保鲜能力,同时混合iturin A,探究得到512 μg/mL iturin A与1%(体积分数)肉桂油复配使用,可显著降低樱桃番茄果实软腐病的发生率,并且可保障番茄的良好贮藏品质。高倩等[64]配合使用生物保鲜技术,混合乳酸链球菌素,也同样具备延长番茄保质期的功能。乙烯对番茄贮藏期的长短也至关重要[87],百里香酚与乙烯清除剂KMnO4联合使用,可更大限度地保持番茄贮藏期的品质[63]。
在番茄的贮藏中,植物提取物保鲜技术无论单独使用还是联合其他保鲜手段,都已经取得成效,但想要进一步完善,需要新想法的引入。袁帅等[88]从代谢入手,通过构建不同代谢网络,测定并计算得到苹果的代谢通量,获得苹果的最佳贮藏温度,可有效延长苹果保质期,防止发生冷害。果蔬的保鲜本质是为了延缓衰老,降低呼吸作用,减少分解代谢,故结合对番茄的代谢研究,可为植物提取物保鲜技术在番茄贮藏上提供新的思路和研究依据。
番茄的营养价值高,产品类型多,副产品也具有很高的利用价值,所以延缓番茄的衰老,增强番茄的货架期尤为重要。在众多保鲜技术中植物提取物保鲜技术更加符合现在人们健康、环保的生活饮食观念,但植物提取物保鲜技术在番茄贮藏上的应用还不够成熟,需要寻求突破,进行更深入地研究,而与不同保鲜技术相结合或是新的保鲜技术的开发或许可以打破此保鲜技术的局限,在一定程度上缓解问题。同时也应注重对微生物、温度等因素的把控,在机理与提取研究方面加大投入力度,才能使植物提取物保鲜技术更好地应用于番茄,乃至果蔬、肉类等领域的保鲜。
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