橙汁腐败酵母菌的分子鉴定及其形态特征分析

doi:10.7506/spkx1002-6630-201209028

食品科学 ›› 2012, Vol. 33 ›› Issue (9): 131-137.doi: 10.7506/spkx1002-6630-201209028

橙汁腐败酵母菌的分子鉴定及其形态特征分析

郭冬琴1，戴木兰1，周志钦1,2，焦必宁2,3，杨晓红1,2,*

1.西南大学园艺园林学院 2.南方山地园艺学教育部重点实验室 3.中国农业科学院柑桔研究所

出版日期:2012-05-15 发布日期:2012-05-07
基金资助:
“十一五”国家科技支撑计划项目(2007BAD47B07-01)；西南大学研究生教育创新计划项目(kb2010012)

Molecular Identification and Morphological Analysis of Yeasts in Spoiled Orange Juice

GUO Dong-qin1，DAI Mu-lan1，ZHOU Zhi-qin1,2，JIAO Bi-ning2,3，YANG Xiao-hong1,2,*

(1. College of Horticulture and Landscape Architecture, Southwest University, Chongqing 400715, China； 2. Key Laboratory of Horticulture Science for Southern Mountainous Regions, Ministry of Education, Chongqing 400715, China； 3. Citrus Research Institute, Chinese Academy of Agricultural Sciences, Chongqing 400712, China)

Online:2012-05-15 Published:2012-05-07

摘要/Abstract

摘要： 目的：结合多种酵母菌鉴定方法，分析腐败橙汁中酵母菌的种类，为快速检测和控制商品橙汁中酵母菌污染奠定基础，也为酵母菌种类的快速鉴定提供参考。方法：以分离自腐败橙汁的8株酵母菌株为材料，观察菌落和细胞形态。用引物ITS1/ITS4扩增菌株5.8S rDNA 及其ITS间隔区(5.8S-ITS)，对扩增产物用HhaⅠ、Hae Ⅲ 和 HinfⅠ进行限制性酶切片段多态性(RFLP)分析和测序；用引物NL1/NL4扩增26S rDNA D1/D2区并测序。结果：菌落特征观察将菌株初分为6组，细胞学观察粗分为3组，分子方法都分成4组。用限制性内切酶HhaⅠ、Hae Ⅲ和HinfⅠ对5.8S-ITS区产物进行 RFLP分析，观察到4种不同的图谱类型。5.8S-ITS 区和26S rDNA D1/D2区的序列分析结果相似，8株分离菌与GenBank中的4种酵母菌参考菌株序列一致性达99%以上。分离株与参考菌株以两区序列构建的NJ系统树都分成4枝：Y1与克鲁维毕赤酵母(Pichia kluyveri)，Y12-3、Y18-1与发酵毕赤酵母(Pichia fermentans)，Y22、Y23、Y26和Y56与Meyerozyma guilliermondii，Y47与Wickerhamomyces anomalus分别聚为一枝。结论：结合形态分析和核酸分析，将8株腐败橙汁分离菌鉴定为P. kluyveri var. kluyveri、P. fermentans、M. guilliermondii和W. anomalus 4个种，其中M. guilliermondii在腐败橙汁中首见报道；ITS- RFLP分析、26S rDNA D1/D2区测序与菌株形态特征结合能有效鉴定酵母菌，核糖体DNA分析可鉴定酵母到生物学种，菌落形态特征可反映种以下的遗传差异，因此，采用分子方法鉴定时不能忽视形态特征分析的重要性。

关键词: 腐败橙汁, 酵母菌, 分子鉴定, 形态分析

Abstract: Objective: To effectively investigate yeasts involved in the spoilage of orange juice and lay the basis for the rapid detection, identification and control of yeasts in juice products. Methods: The colonial and cellular morphology of 8 yeast isolates from spoiled orange juice was observed. The 5.8S rDNA and two ribosomal internal transcribed spacers (5.8S-ITS) of yeasts were amplified with primer ITS1/ITS4. The PCR products were digested with Hha Ⅰ, Hae Ⅲ and Hinf Ⅰ, and then were sequenced using restriction fragment length polymorphism (RFLP). The D1/D2 domain of 26S rDNA was also amplified with primer NL1/NL4 and sequenced. Results: Eight yeast isolates were divided into six groups according to their colonial morphology, three groups according to cellular morphology, and four groups according to molecular analysis. RFLP analysis of 5.8S-ITS region resulted in four restriction patterns. The sequence analysis results of 5.8S-ITS region were similar with those of D1/D2. The sequence identity of eight isolates and four reference strains was as high as 99%. The phylogenetic trees constructed based on 5.8S-ITS region and D1/D2 domain were almost the same and had four branches: isolate Y1 clustered with Pichia kluyveri, Y12-3 and Y18-1 clustered with Pichia fermentans, Y22, Y23, Y26 and Y56 clustered with Meyerozyma guilliermondii, and Y47 clustered with Wickerhamomyces anomalus. Eight yeast isolates from spoiled orange juice were identified as P. kluyveri var. kluyveri, P. fermentans, M. guilliermondii and W. anomalus. M. guilliermondii is observed in spoiled orange juice for the first time. The combination of ITS-RFLP analysis, 26S rRNA gene sequencing and morphological analysis was effective for the identification of spoilage yeasts in orange juice. Therefore, rDNA analysis can allow the identification of yeast species. Colony characteristics may imply genetic differences at various species levels. Furthermore, the importance of morphological analysis cannot be ignored when yeasts are identified by molecular methods.

Key words: spoiled orange juice, yeasts, molecular identification, morphological analysis

中图分类号:

Q93-331

郭冬琴，戴木兰，周志钦，焦必宁，杨晓红. 橙汁腐败酵母菌的分子鉴定及其形态特征分析[J]. 食品科学, 2012, 33(9): 131-137.

GUODong-qin，DAIMu-lan，ZHOUZhi-qin，JIAOBi-ning，YANGXiao-hong. Molecular Identification and Morphological Analysis of Yeasts in Spoiled Orange Juice[J]. FOOD SCIENCE, 2012, 33(9): 131-137.

[1]	牟志勇，杨昳津，王光强，熊智强，宋馨，李国辉，艾连中，夏永军. 酵母菌的益生功能及在食品中的应用[J]. 食品科学, 2021, 42(15): 309-318.
[2]	赵艳芳，康绪明，宁劲松，翟毓秀，尚德荣，丁海燕，盛晓风. 虾蛄可食组织中镉和砷的形态及分布特征[J]. 食品科学, 2020, 41(8): 282-287.
[3]	罗方雯，黄永光，涂华彬，胡峰，尤小龙. 基于高通量测序技术对茅台镇酱香白酒主酿区域酵母菌群结构多样性的解析[J]. 食品科学, 2020, 41(20): 127-133.
[4]	向慧平，林宜锦，关统伟，张家旭，尚红光，赵小林，焦士蓉，冯栩，杨阳. 四川浓香型大曲生产中酵母菌、芽孢杆菌与工艺指标的关联性分析[J]. 食品科学, 2020, 41(2): 196-201.
[5]	张宁，范迪，杨燕，王赢，徐琼，崔学文，房玉国，闻宏亮，秦峰，刘浩，范一灵. 食品中肌醇测定用菌株的筛选及检测方法建立和应用[J]. 食品科学, 2020, 41(10): 88-93.
[6]	吴丹丽，李高磊，王明月，陈晓涵，李余动. 低产尿素与高级醇黄酒酵母菌株的筛选、鉴定与发酵[J]. 食品科学, 2019, 40(6): 100-105.
[7]	赵鲁宁，周秋阳，杨慧慧，郑鸿儒，温新宇，孙卫红. 季也蒙毕赤酵母Y35-1菌株对枇杷采后炭疽病的抑菌效果及保鲜作用[J]. 食品科学, 2019, 40(4): 170-177.
[8]	李丽杰，贺敏，贺银凤，孙禹. 酵母菌对重金属的吸附与抗性和解毒重金属的胞内分子机制研究进展[J]. 食品科学, 2019, 40(3): 330-337.
[9]	赵英杰，张文平，吴剑梅，程新. 乳扇中高产胞外多糖酵母菌的筛选及其抗氧化活性[J]. 食品科学, 2019, 40(22): 134-140.
[10]	唐庆强，曹晓钢，夏林兵，陈锦丽，张凤珍，罗顺玉，王丹红,. As(Ⅲ)和As(Ⅴ)在香菇栽培基质和子实体中的迁移规律[J]. 食品科学, 2019, 40(21): 94-99.
[11]	陈贵宇，潘煜辰，李清清，冷桃花，李君绩，葛宇. 高效液相色谱-电感耦合等离子质谱法分析富硒茶叶中硒的形态[J]. 食品科学, 2018, 39(8): 155-159.
[12]	姚博，贠建民，艾对元，严海娇，白杰，李多佳. 浆水中产香酵母菌菌株的筛选及其增殖培养基优化[J]. 食品科学, 2018, 39(6): 107-115.
[13]	龙兴瑶，李键，谢婕，赵欣，王洪伟，张玉，索化夷,. 藏灵菇牦牛酸乳发酵过程中乳酸菌和酵母菌对品质的作用[J]. 食品科学, 2018, 39(4): 82-89.
[14]	陈尚龙，刘恩岐，陈安徽，刘辉，巫永华，秦旭. 应用体外全仿生模型初步分析2 种富硒产品中硒形态及生物可给性[J]. 食品科学, 2018, 39(4): 225-232.
[15]	文淑婷，李艳. 老白干酒曲中酵母菌多样性分析[J]. 食品科学, 2018, 39(24): 175-182.

橙汁腐败酵母菌的分子鉴定及其形态特征分析

Molecular Identification and Morphological Analysis of Yeasts in Spoiled Orange Juice

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价