冷冻保护剂和储藏条件对海洋酵母冻干粉活性的影响

海洋拮抗酵母Rhodosporidium paludigenum作为生防保鲜剂在控制水果采后真菌类病原菌中表现出了良好的抑制效果,然而生防制剂要真正取代化学杀菌剂,必须以产品的形式投入市场,即实现其商品化生产。实验将该拮抗酵母制成冻干粉,研究冻干保护剂和储藏条件对海洋酵母干粉稳定性的影响。结果显示,海藻糖作为保护剂可以显著地提高酵母干粉的存活率和生防活性,用5%的海藻糖作保护剂,存活率为42%,而对照仅为3%。研究还发现,酵母细胞的存活率与其胞内海藻糖含量具有一定的相关性。另外,储藏实验表明,低温(4℃)可以较好地保持酵母干粉的存活力。而常温(25℃)下储藏1个月,每克干粉中酵母活细胞数下降5个数量级,因此常温不适合储藏该新型拮抗酵母生防制剂。     

酵母甘露聚糖处理对番茄果实贮藏效果的影响

为了研究酵母甘露聚糖对番茄果实贮藏效果的影响,利用不同质量浓度（0、1.0、10.0 g/L）酵母甘露聚糖溶液对番茄果实进行负压渗透处理,测定贮藏过程中果实硬度、可溶性固形物含量、色差、转色指数、乙烯释放速率、呼吸强度、β-半乳糖苷酶活性及损伤接种链格孢菌等指标。结果表明：酵母甘露聚糖能够延缓番茄果实硬度的下降,减缓可溶性固形物含量的变化,降低乙烯释放速率和呼吸强度,推迟果实转色,从而有效地延缓了番茄果实后熟进程,提高了贮藏效果。其中,1.0 g/L酵母甘露聚糖作用效果最为显著。进一步测定表明,酵母甘露聚糖处理可抑制番茄果实β-半乳糖苷酶活性变化。损伤接种链格孢菌实验表明,酵母甘露聚糖还能有效地抑制由链格孢菌引起的番茄果实黑霉病,减小病斑面积,降低病害发病率。     

Biological control of postharvest green mold decay of oranges by Rhodotorula glutinis

The biocontrol activity of Rhodotorula glutinis on green mold decay of oranges caused by Penicillium digitatum was investigated in vitro and in vivo. Significant control was achieved with a washed cell suspension and an unwashed cell culture mixture of R. glutinis. Treatment of wounds with autoclaved cell cultures or cell-free culture filtrate did not prevent decay. The protection provided by the washed yeast cells was dose-dependent. The higher the concentration of R. glutinis, the better the effect of the biocontrol capacity. At concentrations of yeast of 1×10⁹ colony-forming units per milliliter or higher and pathogen spore suspensions of 5×10⁴ spores per milliliter, green mold was almost inhibited after 4-days incubation at 20 °C. The interval between the pathogen inoculation and the antagonist application significantly influenced the biocontrol ability. The biocontrol efficacy of R. glutinis applied before the pathogen was better than that of applied after the pathogen. Surprisingly, R. glutinis was also effective in controlling green mold at low temperature (4 °C). Rapid colonization of the yeast in wounds was observed during the first 3 days at 20 °C, and remained stable after 5-days incubation. On fruits stored at 4 °C, even after 21 days, the population of R. glutinis in wounded fruits was more than 1,600-fold of what it was just prior to storage. In the test on potato dextrose agar plates, agar disks of R. glutinis nutrient yeast dextrose agar cultures placed on PDA plates seeded with pathogens did not inhibit the growth of P. digitatum. Spore germination of pathogens in potato dextrose broth was greatly controlled in the presence of living cell suspensions.     

L-谷氨酸对樱桃番茄果实病程相关蛋白的影响

为研究L-谷氨酸(谷氨酸)对果实采后病害的抗性机制,以樱桃番茄为对象,将其浸泡于100 mg/L谷氨酸溶液10 min,经不同诱导时间(0、12、24、36 h)处理后接种Alternaria alternata,定期测定果实的发病率,同时研究谷氨酸对果实病程相关蛋白(pathogenesis-related proteins,PRs)及其编码基因的影响。在体外条件下,观察谷氨酸对病原菌生长状态的影响。结果表明:谷氨酸诱导时间在24 h及以上时,番茄发病率显著下降;体外试验中,谷氨酸对病原菌的孢子萌发率和生长无明显影响;谷氨酸可迅速提高果实β-1,3-葡聚糖酶、几丁质酶(chitinases,CHI)和过氧化物酶(peroxidase,POD)的活力,且诱导LePR1、LePR5、LeCHI3、LeCHI9和LePOD上调表达。综上,谷氨酸对Alternaria alternata的作用机制可能与其诱导了果实体内的病程相关蛋白有关。     

Control of postharvest pear diseases using Rhodotorula glutinis and its effects on postharvest quality parameters

Rhodotorula glutinis was evaluated for its activity in reducing postharvest gray mold decay and blue mold decay of pear caused by Botrytis cinerea and Penicillium expansum respectively, and in reducing natural decay development of pear fruits, as well as its effects on postharvest quality of fruits. There was a significant negative correlation between concentrations of the yeast cells and infectivity of the pathogens. At concentrations of R. glutinis at 5x108 CFU/ml, the gray mold decay was completely inhibited after 7 days incubation at 20 degrees C, while the control fruit had 100% disease incidence and 2.15 cm lesion diameter respectively, at challenged with B. cinerea spores suspension of 1x105 spores/ml; No completely control was got to blue mold, when pear fruits stored at 20 degrees C for 7 d (challenged with P. expansum spores suspension of 5x104 spores/ml), but the decay was distinctly prevented with 20% and 0.60 cm of disease incidence and lesion diameter respectively, while the control fruits were 100% and 2.74 cm, respectively. Rapid colonization of the yeast in wounds was observed during the first 1 d at 20 degrees C, and then the populations stabilized for the remaining storage period. On pear wounds kept at 4 degrees C, rapid colonization of the yeast in wounds was observed during the first 3 d, and then the increase in population density of R. glutinis turned slow, which continued over 6 d after application of the antagonist until it reached a high level. Then, the populations stabilized for the remaining storage period. In the test on PDA plates, R. glutinis significantly inhibit the growth of B. cinerea and P. expansum. Spore germination of pathogens in PDB was greatly controlled in the present of living yeast cell suspensions. R. glutinis significantly reduced the natural development of decay of pear following storage at 20 degrees C for 7 days or at 4 degrees C for 30 days followed by 20 degrees C for 5 days, and did not impair quality parameters, including mass loss, firmness, TSS, ascorbic acid or titratable acidity. Thus, the application of R. glutinis can be an alternative to chemicals for control of postharvest diseases on pear fruits.     

ε-聚赖氨酸对樱桃采后病原菌橘青霉和链格孢菌的抑菌作用研究

目的 探究ε-聚赖氨酸(ε-polylysine,ε-PL)对樱桃果实采后病原菌橘青霉(Penicillium citrinum)和链格孢菌(Alternaria alternata)的抑菌作用和抑菌机制.方法 本研究测定100、200、400、800 mg/Lε-PL处理对2种病原菌菌丝生长、孢子萌发、细胞膜透性的影...     

Synergistic effects of combining biocontrol agents with silicon against postharvest diseases of jujube fruit

The synergistic effects of biocontrol yeasts Cryptococcus laurentii and Rhodotorula glutinis combined with silicon (Si) against Alternaria alternata and Penicillium expansum molds were investigated in jujube fruit (Chinese date, Zizyphus jujuba) stored at 20 and 0 degrees C, respectively. Combinations of C. laurentii and R. glutinis at 5 x 10(7) cells/ml with 2% Si was most effective in controlling the diseases caused by A. alternata and P. expansum on jujube fruit stored at 20 degrees C. When fruits were stored at 0 degrees C, combining C. laurentii and R. glutinis with Si was as effective against P. expansum as was Si or the yeasts applied alone and was more effective in controlling A. alternata. Si may have a fungistatic effect by directly inhibiting pathogen growth, but it did not greatly influence the growth of the antagonists.     

Effect of water activity on stress tolerance and biocontrol activity in antagonistic yeast Rhodosporidium paludigenum

The aims of this study were to optimize the salt-adaptation conditions of the marine antagonistic yeast Rhodosporidium paludigenum and investigate the biocontrol activity of salt-induced cell suspensions of R. paludigenum on postharvest pathogens in fruits. Low water activity (a_w = 0.98, 0.97, 0.96, and 0.95) inhibited the growth of R. paludigenum in nutrient yeast dextrose broth, but the yeast grew better in the medium modified with NaCl solute than other nonionic solutes. R. paludigenum grown in 6.6% NaCl-modified medium had higher viabilities (92.1%) at low water activity (a_w = 0.95) than control (81.1%) after 48 h incubation. The salt-adapted R. paludigenum also showed better viability than the un-adapted cells after being frozen, which may be related to the accumulation of intracellular trehalose. Moreover, the best biocontrol inhibition in pears and Chinese winter jujubes was obtained when R. paludigenum was grown in NaCl-modified medium. Therefore, this study implies that improving physiological inducement methods may be a promising strategy for accelerating commercialization of biocontrol agents.     

Effect of gamma irradiation and water activity on mycotoxin production of Alternaria in tomato paste and juice

Gamma-irradiation, water activity (aw) and incubation temperature were found to affect the production of tenuazonic acid (TZA) by Alternaria alternata in tomato paste and juice. By increasing the irradiation doses, the dry weight as well as TZA decreased greatly until complete inhibition at 4 kGy. Greatest production of TZA occurred at 0.98 aw (57.5 micrograms/g and 26.3 micrograms/g) for both tomato paste and juice, respectively, at 25 degrees C. Changing temperature and aw altered the relative amounts of TZA produced in tomato paste und juice by unirradiated and irradiated conidia of A. alternata. Only trace amount of TZA was detected at 0.98 aw (1.50 micrograms/g) by 3 kGy-irradiated conidia in tomato paste, while it was inhibited completely in juice. Increasing gamma-irradiation doses and decreasing water activities decreased greatly or inhibited TZA production in both tomato paste and juice.     

苹果腐败霉菌的拮抗酵母菌筛选鉴定及拮抗机理初探

从苹果果皮中筛选出对引起苹果采后病害的扩展青霉(Penicillium expansum)、互隔交链格孢霉(Alternaria alternata)和黑曲霉(Aspergillus niger)具有拮抗作用的酵母菌,并初步研究其拮抗机理。结果表明:筛选得到两株可以同时拮抗三种腐败霉菌的拮抗酵母菌Y4和Y9,根据酵母菌菌落性状、生理生化特征并结合分子生物学的分析结果,鉴定为Candida railenensis和Aureobasidium pullulans,其中A.pullulans为类酵母真菌。C.railenensis的最佳拮抗浓度为109 CFU/mL,对P.expansum、A.alternata和A.niger的菌丝生长、孢子萌发和芽管生长均有显著(p<0.05)的抑制作用。扫描电镜观察到C.railenensis能够包围和吸附腐败霉菌孢子,与它们竞争营养与空间,此外还能够吸附P.expansum菌丝体与其直接作用。C.railenensis的主要抑病机理是营养与空间竞争,并且其生防效果的发挥是多种方式共同作用的综合结果。     

生防菌KD-16J的鉴定及其对番茄灰霉病的抑制作用

通过离体和活体筛选,确定一株生防菌KD-16J对果实灰霉病的抑制效果。经形态学鉴定、生理生化试验和内源转录间隔区序列分析鉴定,确定其为膜醭毕赤酵母菌(Pichia membranifaciens)属。在平板抑菌试验中发现,在105 CFU/mL~108 CFU/mL之间,酵母菌的应用浓度越高,对灰葡萄孢菌的抑制效果越好,1×108 CFU/mL的生防菌对灰葡萄孢菌菌丝生长的抑制率为85.53%,同时能够完全抑制孢子的萌发;显微镜观察发现与酵母菌共培养能够导致灰葡萄孢菌菌丝肿胀,内部物质泄漏。活体试验(番茄果实)结果表明,该菌株能够显著抑制番茄果实的自然发病率和有伤接种灰葡萄孢菌果实的灰霉病发病率,而且喷洒该菌的时间越早对果实灰霉病的抑制效果越好,当生防菌KD-16J先于灰霉菌36 h处理时,番茄果实的发病率仅为29.12%。     

采前3 种杀菌剂喷施对樱桃番茄采后品质的影响及啶菌噁唑残留检测

以金珠樱桃番茄为试材,研究采前杀菌剂喷施对采后樱桃番茄发病率、质量损失率和食用品质的影响,并测定樱桃番茄果实中啶菌噁唑残留变化。结果表明：采前喷施杀菌剂显著降低了金珠樱桃番茄采后发病率,提高了樱桃番茄果实中VC和可溶性固形物含量,减缓了VC和可溶性固形物含量贮藏期间的损失;但对质量损失率和可滴定酸含量的影响不显著（P＜0.05）。对樱桃番茄采后贮藏保鲜效果最好的杀菌剂为啶菌噁唑,喷施质量浓度为0.4 g/L。采前喷施啶菌噁唑后,樱桃番茄中啶菌噁唑残留在采前和采后均呈下降趋势;且采后樱桃番茄中啶菌噁唑残留下降的速度比采前下降的慢,贮藏温度越低,下降越慢。     

施硒对樱桃番茄贮藏品质及3种活性氧代谢酶的影响

为探寻施硒对樱桃番茄采后贮藏品质的影响并确定其适宜的处理浓度,在樱桃番茄发育过程中分次冲施不同浓度硒肥,分析番茄果实采收时及在贮藏过程中果实硬度、色泽、乙烯释放量、糖、酸、V_C含量及活性氧代谢相关酶活性的变化情况。结果表明,采收时2 mg/L硒处理的樱桃番茄果实硬度、可溶性糖含量、V_C含量与对照组相比分别增加了7.98%、5.29%和11.32%,并在贮藏过程中始终维持较高含量;同时2 mg/L硒处理提高了采后果实过氧化物酶活性(40.01~64.24 U/g)和超氧化物歧化酶活性(36.94~186.39 U/g);并在乙烯释放高峰时期(21 d)抑制乙烯释放量至8.01 μL/kg·h,延缓了樱桃番茄果实的采后衰老。而3 mg/L硒处理在贮藏42 d时对果实的活性氧代谢酶表现出负面影响,过氧化氢酶、过氧化物酶、超氧化物歧化酶活性分别低于对照组11.93%、23.09%和31.01%,不利于维持番茄果实的贮藏品质。2 mg/L为适宜的采前硒处理浓度,可以提升樱桃番茄果实品质及贮藏特性。     

海洋红冬孢酵母分泌蛋白的鉴定及其在番茄采后生物防治中的应用

以番茄果实为实验试材，选取海洋红冬孢酵母（Rhodosporidium paludigenum）为研究对象，利用Q Exactive UHMR组合型四极杆Orbitrap质谱鉴定并结合生物信息学分析其分泌蛋白组成，探究分泌蛋白在微生物之间以及微生物与番茄之间建立互作关系的功能机制。结果表明，R. paludigenum发酵上清液中含有1 632 条红酵母（Rhodotorula）源特异性目标蛋白质多肽，结合Signal P、WoLF PSORT、Target P、TMHMM、big-PI Predictor和Secretome P等软件分析得到58 个经典分泌特征的蛋白质和433 个非典型分泌特征的蛋白质。蛋白生物学功能结果表明，R. paludigenum分泌蛋白可有效抑制灰葡萄孢菌孢子萌发，抑制率为31.4%，同时有效增强番茄果实伤口处的系统抗病能力，抑制果实伤口处灰霉病发病几率，抑制率达50.1%。结合UniParc数据库信息，发现R. paludigenum分泌蛋白中含有藻胶裂解酶和β-葡萄糖苷酶等18 个碳水化合物酶类蛋白（31.0%），推测可能在R. paludigenum与灰葡萄孢菌的互作和诱导番茄果实系统抗性过程中发挥重要功能，以保护番茄果实免受采后灰霉病害的侵染。     

Survival of Salmonella Montevideo on tomato leaves and mature green tomatoes

Survival of Salmonella Montevideo on tomato leaf and mature green tomatoes was examined. When spiked on the surfaces of excised leaves from greenhouse tomato plants, Salmonella Montevideo survived poorly if the leaves were allowed to dry. After spot inoculation of 6.6 log CFU per leaflet and incubation for 48 h at 60% relative humidity, the leaflets were dried, and surviving Salmonella were reduced 2.8 to 3.7 log CFU per leaflet. However, when leaves spiked with Salmonella were supported on hydroponic nutrient medium and incubated at 100% relative humidity, there was no significant reduction of surviving bacteria for at least 6 days. Exposure of mature green tomatoes to ethylene (100 ppm at 100% relative humidity and 20 degrees C for 6 days) did not significantly affect the survival of Salmonella on their surfaces. The significance of these data to pre- and postharvest safety of tomato is discussed.     

1-甲基环丙烯处理对樱桃番茄果实低温贮藏品质的影响

为了探讨低温贮藏条件下1-甲基环丙烯(1-methylcyclopropene,1-MCP)处理对樱桃番茄果实采后品质的影响,该研究以'粉娇'樱桃番茄为试材,测定了经1-MCP(0.14μL/L)处理的樱桃番茄在低温[(8±1)℃]贮藏期间果实腐烂率、呼吸强度、果实硬度、可溶性固形物、维生素C、有机酸和类胡萝卜素含量的...     

A Box-Behnken design for predicting the combined effects of relative humidity and temperature on antagonistic yeast population density at the surface of apples

The objective of this work was to develop models predicting the combined effects of relative humidity (RH, 75-98%), temperature (5-25 degrees C), and initial applied yeast concentration (10(4)-10(8) CFU/ml) on the apple-surface population densities of two biocontrol agents fused against postharvest diseases; the antagonistic yeasts Pichia anomala strain K and Candida oleophila strain O. Experiments were carried out according to a Box-Behnken matrix. Multiple regression analyses showed that both models yielded a good prediction of yeast density. The effect of relative humidity appeared greater than that of temperature. The number of yeast colony-forming units per square centimeter of apple fruit surface increased with increasing relative humidity, temperature, and initial applied yeast concentration. The models predict that under optimal growth conditions (25 degrees C, 98%), strains O and K should reach a density of 10(4) CFU/cm2 when applied initially at 2 x 10(7) (strain O) or 10(7) CFU/ml (strain K). The model results suggest that rainfall was likely the principal cause of the variability of yeast efficacy reported for previous preharvest orchard trials spanning two successive years. Temperature may also contribute to this variation. The models developed here are important tools for predicting population densities of both strains on the apple surface within the experimental limits. The use of these results should contribute to achieving yeast densities of 10(4) CFU/cm2 on apples by controlling yeast application and environmental factors such as relative humidity and temperature. The results of this study also confirm our previous in vitro findings that water activity has a greater effect than temperature on yeast population density.     

吡啶-2,6-二羧酸处理对梨果采后黑斑病控制作用及其抑菌机理

由互隔交链孢（Alternaria alternata）引起的黑斑病是梨果主要采后病害之一,会导致严重的经济损失。本实验以梨果黑斑病菌A. alternata为研究对象,通过体内外实验研究了枯草芽孢杆菌（Bacillus subtilis）代谢产物吡啶-2,6-二羧酸（pyridine-2,6-dipicolinic acid,DPA）对A. alternata生长的抑制和对梨果黑斑病的控制作用,并进一步探讨了其抑菌作用的机理。结果表明：DPA能显著抑制A. alternata的菌落生长、孢子萌发以及芽管伸长,其中2.0 mmol/L DPA处理后培养9 d时A. alternata菌落直径仅为对照组的51.76%,4.0 mmol/L处理可完全抑制A. alternata的生长。DPA处理对梨果黑斑病的扩展有明显的控制作用,贮存至第11天时,20.0 mmol/L DPA处理组早酥梨病斑直径仅为对照组的55.17%。进一步研究表明,DPA处理提高了A. alternata细胞膜的通透性,菌丝电导率、蛋白质和核酸渗漏量显著增加,同时DPA处理降低了细胞膜组分麦角甾醇质量浓度,提高了A. alternata丙二醛浓度。结论：DPA可通过延缓A. alternata生长、提高膜通透性、破坏细胞膜的完整性和促使膜脂氧化而发挥抑菌作用。     

超声协同钙浸渍对樱桃番茄贮藏过程中酚类物质的影响研究

研究超声协同钙浸渍处理对樱桃番茄贮藏过程中酚类物质的含量、结构和抗氧化能力以及相关酶活性的影响。结果表明:超声协同钙浸渍处理有效抑制贮藏期间樱桃番茄果实中多酚氧化酶(polyphenol oxidase,PPO)、过氧化物酶(peroxidase,POD)活性和苯丙氨酸转氨酶(phenylalanine ammonia-lyase,PAL)活性,维持总酚含量,延缓后熟。质谱结果表明,超声协同钙浸渍技术有效抑制贮藏期间樱桃番茄果实主要酚酸的消耗,更好的维持酚类物质的结构特性和抗氧化活性。综上所述,超声协同钙浸渍技术能够有效维持樱桃番茄贮藏期间品质特性,增加果实耐贮藏性。     

Efficacy of preharvest and postharvest Candida sake biocontrol treatments to prevent blue mould on apples during cold storage

This study compared the biocontrol efficiency of preharvest and postharvest applied yeast cells of Candida sake to apples (cv. Golden Delicious) wounded before and after harvest and inoculated with Penicillium expansum prior to cold storage conditions in two seasons, 1994/95 and 1995/96. The establishment of populations of C. sake during this period also was determined. In both years, postharvest treatment with the antagonist resulted in significant (P<0.05) and effective control of Penicillium rot whether pre- or postharvest wounds were made. Maximum disease control achieved, in terms of incidence and severity, was greater than 80% reduction in lesion diameter and 50% reduction in the incidence of lesions. However, preharvest application of the antagonistic yeast at a concentration of 3·10⁶ CFU/ml was less effective against Penicillium rot than postharvest treatment. No advantages in biocontrol were observed when apples were treated with the yeast antagonist both pre- and postharvest. Candida sake population levels during cold storage of apples receiving only preharvest application of the antagonist decreased prior and more rapidly than levels in apples receiving a postharvest application. High populations of C. sake were present in postharvest-treated apples, even after 90 days in cold storage.