雷州半岛近岸海域海洋红酵母的分离鉴定
作者简介:孙建男(1989—), 女, 辽宁省朝阳市人, 硕士, 研究方向为微生物学与分子生物学。E-mail:1042341150@qq.com
收稿日期: 2016-11-16
要求修回日期: 2016-12-27
网络出版日期: 2017-07-26
基金资助
广东省科技计划项目(2013B020307013)
广东省农业科技项目(201201147)
湛江市科技计划项目(2015B01013)
Isolation and identification of marine Rhodotorula in a coastal area near Leizhou Peninsula
Received date: 2016-11-16
Request revised date: 2016-12-27
Online published: 2017-07-26
Supported by
Science and Technology Program of Guangdong Province (2013B020307013)
Agricultural Science and Technology Program of Guangdong Province (201201147)
Science and Technology Program of Zhanjiang (2015B01013)
Copyright
从雷州半岛近岸海域采集海洋生物、海水、海泥等样品, 共计317份。将样品处理后先富集培养, 再用红酵母琼脂培养基及麦芽汁琼脂培养基进行分离培养, 之后对分离菌株通过菌体形态、培养特性及生理生化试验结果进行初步鉴定。根据红酵母的初步鉴定结果, 选取有代表性的菌株作26s rDNA D1/D2区域序列测定, 构建系统发育树, 进行多样性分析。结果表明, 从317份样品中分离鉴定出71株海洋红酵母; 其中, 有代表性的36株红酵母归属于5个种和2个疑似新种。36株红酵母包括胶红酵母(Rhodotorula mucilaginosa)22株、粘红酵母(Rhodotorula glutinis)6株、红冬孢酵母(Rhodosporidium toruloides)3株、斯鲁菲亚红酵母(Rhodotorula slooffiae)2株、浅红红酵母1株(Rhodotorula pallida)和疑似红酵母新种2株(Rhodotorula sp, cZDJ2和Rhodotorula sp. ZTC2)。雷州半岛近岸海域的海洋红酵母优势种为胶红酵母, 种群的多样性不够丰富。
孙建男 , 刘影 , 谢为天 , 徐春厚 . 雷州半岛近岸海域海洋红酵母的分离鉴定[J]. 热带海洋学报, 2017 , 36(4) : 87 -92 . DOI: 10.11978/2016117
In this study, 317 samples from marine organisms, seawater and sea mud were gathered in a coastal area near the Leizhou Peninsula. After basic process and enrichment culture, marine Rhodotorula was isolated by using Rhodotorula agar medium and wort agar medium. The Rhodotorula strains were preliminary identified by using cell morphology characteristics, cultural characteristics, physiological and biochemical test results. Based on the preliminary identification results, representative strains were tested by 26s rDNA D1/D2 domain sequence analysis, phylogenetic tree was constructed and diversity was analyzed. The 71 strains isolated from 317 samples were preliminary identified as marine Rhodotorula among which 36 representative strains were identified as belong to five species and two suspected to be new species. Among them, 22 strains were R. mucilaginosa, six strains were R. glutinis, three strains were R. toruloides, two strains were R. slooffiae, one strain was R. pallida, and two strains were suspected to be new species named Rhodotorula sp. cZDJ2 and Rhodotorula sp. ZTC2, respectively. The dominant species of marine Rhodotorula in the coastal area near the Leizhou Peninsula was Rhodotorula mucilaginosa, indicating that the diversity of population in the coastal area near the Leizhou Peninsula was not rich enough.
Fig. 1 Colony morphology of Suspected Rhodotorula strains图1 疑似红酵母菌株的菌落形态 |
Fig. 2 Cell morphology of Suspected Rhodotorula strains (×1000)图2 疑似红酵母菌株的菌体形态(×1000) |
Tab.1 Assimilation tests of carbon and nitrogen in 71 strains表1 71株分离菌的碳源和氮源同化生长试验 |
分离菌株 | 蔗糖 | 麦芽糖 | 海藻糖 | 松三糖 | 木糖 | 鼠李糖 | 乳糖 | 蜜二糖 | 棉子糖 | 甘露糖 | 菊糖 | 肌醇 | 淀粉 | 硝酸盐 | 亚硝 酸盐 | 乙胺 | 肌酸 | 肌酸酐 |
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
ZTC4等46株 | + | + | + | + | + | v | - | - | + | v | - | - | - | - | - | + | - | - |
aZDD4等12株 | + | + | + | + | v | + | - | - | + | + | + | - | - | + | + | + | - | - |
ZTHD2等6株 | + | + | + | + | + | - | - | - | v | v | v | - | - | + | + | v | - | - |
aZDB等4株 | + | - | + | + | + | v | + | - | v | v | v | - | - | + | + | - | - | - |
ZZC2 | + | - | + | - | - | - | - | - | - | + | - | - | - | - | - | - | - | - |
ZTC2 | - | + | + | - | + | + | - | - | + | + | - | - | - | + | + | + | - | - |
cZDJ2 | + | - | + | + | + | - | - | - | - | + | - | - | - | - | - | - | - | - |
注: “+”表示结果阳性; “-”表示结果阴性; “v”表示结果不确定; 空白表示无数据 |
Tab. 2 26S rDNA D1/D2 sequence similarity between 36 Rhodotorula strains and their relationships most closely type strain表2 36株红酵母与其关系最密切的模式菌株间的26S rDNA D1/D2序列相似性 |
类群 | 分离菌株(Genbank登录号) | 相似性最高模式菌株 | 相似性/% |
---|---|---|---|
R. mucilaginous (22株) | ZZC1(KP979796)、aZDD6(KP954615) | R. mucilaginosa ATCC 32763 | 99.6 |
ZTHD4(KP954623、ZTHG1(KP954624) | |||
aZDY(KP966487)、ZTHY5(KP966488) | |||
ZTHQ(KP954629) | |||
cZDL1(KP954617)、aZDY1(KP954618) | R. mucilaginosa ATCC 32763 | 99.5 | |
aZDZ1(KP954619)、cZDY5(KP954620) | |||
bZDB2(KP966490)、 ZDS(KP954630) | |||
ZTHY1(KP954625) | R. mucilaginosa ATCC 32763 | 99.3 | |
aZDD3(KP979795) | R. mucilaginosa ATCC 32763 | 99.1 | |
cZDY2(KP954616)、ZTHD3(KP954622) | R. mucilaginosa CBS 8383 | 99.6 | |
ZTC4(KP966489) | R. mucilaginosa CBS 8383 | 99.5 | |
ZTHY4(KP954626)、ZTC3(KP954627) ZTC5(KP954628) 、ZTHY2(KP966486) | R. mucilaginosa KCTC 2829 | 99.6 | |
R. glutinis(6株) | cZDD5(KP979800)、 bZDD5(KP979801) bZDD4(KP979802) 、bZDD1(KP979803) aZDD5(KP979804)、aZDD4(KP979805) | R. glutinis ATCC 32765 | 99.3 |
R. toruloides(3株) | ZTHD2(KP979806)、ZTC1(KP979807) | R. toruloides KCTC 17083 | 99.5 |
ZTHD1(KP979808) | 99.1 | ||
R. slooffiae(2株) | aZDH8(KP979798)、aZDB(KP979799) | R. slooffiae CBS 5706 | 99.5 |
R . pallid(1株) | ZZC2(KP979797) | R. pallid CBS 320 | 99.3 |
Rhodotorula sp.(1株) | cZDJ2(KP979809) | R. minuta JCM 5951 | 93.0 |
Rhodotorula sp.(1株) | ZTC2(KP954620) | R. mucilaginosa ATCC 32763 | 98.7 |
Fig. 3 Neighbor-Joining phylogenetic tree based on 26S rDNA D1/D2 domain sequence analysis图3 基于26S rDNAD1/D2区域序列构建的系统发育树 |
The authors have declared that no competing interests exist.
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